P.1 

OAR Box 1846 

Prepped by Ollie Stewart 

Document Number: 

8) ll-B-5 

Docket Number: 

A-90-07 

Printed 11/18/2008 7:30:14 AM Sheet 8 of 93 
P.2 

^

£ 0 S,<* 

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY 

WASHINGTON, D.C. 20460 

{] t g n »j.m 

FEB 2 5 1992 

MEMORANDUM 

EPA AIR DOCKET 

SUBJECT: Cost Estimates for TSCA Test Guidelines 

FROM: Dan Axelradjl/j 

Regulatory

 cTmpacts Branch 

Economics and Technology Division (TS-779) 

TO: Sheri Dunatchik 

Office of Mobile Sources 

OFFICE OF 

PESTICIDES AND TOXIC 

SUBSTANCES 

Attached, as requested, are cost estimates for the TSCA test 

guidelines being used by your office. This package is divided 

into three sections: 

1. The cost estimates, summarized in a one page table, followed 

by a set detailed estimates for each test guideline; 

2. An explanation of the methodology used to develop the cost 

estimates; and 

3. The resume of Vincent J. Piccirillo, the consultant who 

prepares the cost estimates. 

Please call me at 382-3713 if you have any questions 

concerning this package. 

Attachment 

Printed on Recycled Paper 
TSCA HEALTH EFFECTS GUIDELINES 

P.3 

PROTOCOL / PROTOCOL NUM. SPECIES ROUTE OF ADMIN. 

Acute Inhalation Toxicity 

798.1150 Rats inhalation 

Subchronic Inhalation Toxicity 

798.2450 Rats inhalation 

Combined Chronic Toxicity/Oncogenicity 

798.3320 Rats inhalation 

Inhalation Developmental Toxicity (2 species) 

798.4350 Mice inhalation 

Rats inhalation 

Developmental Toxicity Preliminary Screen 

798.4420 Mice inhalation 

Rats inhalation 

DATE 

06/24/91 

07/19/90 

08/15/90 

07/19/90 

07/19/90 

07/19/90 

07/19/90 

Reproduction/fertility Effects (2-Generation Reproduction) 

798.4700 Rats inhalation 07/19/90 

Salmonella typhimurium Reverse Mutation Assay (Ames Test) 

798.5265 Salmonella typhimurium 08/15/90 

Detection of Gene Mutations in Somatic Cells in Culture 

798.5300 CHO/HGPRT 

In Vitro Mammalian Cytogenetics (Chromo Aber) 

798.5375 CHO Cells 

08/15/90 

08/15/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: Chromosomal Ana 

798.5385 Rats inhalation 07/20/90 

BEST 

ESTIMATE 

11280 

139710 

1267660 

48600 

48730 

23810 

23570 

439650 

3840 

9240 

8660 

lysis (Chromo 

26730 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: Micronucleus Assay 

798.5395 Mice inhalation 07/20/90 24620 

Unscheduled DNA Synthesis in Mammalian Cells in Culture 

798.5550 Rat Hepatocytes 

In Vitro Sister Chromatid Exchange Assay 

798.5900 CHO 

In Vivo Sister Chromatid Exchange Assay 

798.5915 Rats inhalation 

Functional Observation Battery 

798.6050 ACUTE 

Rats inhalation 

SUBCHRONIC 

Rats inhalation 

General Metabolism 

798.7100 Rats inhalation 

06/24/91 

06/24/91 

06/25/91 

07/19/90 

06/08/90 

08/16/90 

10500 

10740 

15870 

15800 

104330 

162450 

TEST COST 

9400 -

103870 • 

968150 • 

40010 -

40190 -

19350 • 

19210 • 

326440 -

2740 • 

7190 • 

6710 -

Aber) 

22390 -

20520 • 

8720 -

8920 • 

13240 -

13200 -

73610 -

135430 -

RANGE 

13340 

177510 

1585950 

58030 

58040 

28660 

28290 

559390 

4970 

11440 

10760 

31470 

29140 

12460 

12760 

18780 

18660 

136500 

192250 

BURDEN 

158 

2046 

21223 

868 

930 

376 

354 

6670 

36 

97 

100 

374 

366 

115 

116 

205 

247 

1666 

3244 
T PROTOCOL #: 798.1150 

Acute Inhalation Toxicity 

P.4 

STUDY DESIGN: 

Group 

l 

2 

3 

Dose Level 

Low Dose 

Mid Dose 

High Dose 

Number of Animals 

5 males, 5 females 

5 males, 5 females 

5 males, 5 females 

TEST LIMIT: 

If a test at a dose concentration of at least 5 

mg/liter or at the maximal achievable concentration, 

with an exposure of 4 hours or more produces no 

compound-related mortality, then a three dose level 

study is not necessary. 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats. 

AGE OF ANIMALS: 

Young adult animals. 

DURATION AND FREQUENCY OF EXPOSURE: 

A single exposure over a period of at least 4 hours 

with animal observations for at least 14 days after 

exposure. 

ROUTE OF EXPOSURE: 

Inhalation, for at least 4 hours. 

BODY WEIGHT: 

All animals prior to dosing, weekly thereafter and at 

death or sacrifice. 

OBSERVATIONS AND PHYSICAL EXAMINATIONS: 

At least cnce daily. More frequent observations should 

be made during the first few days to assure the 

observation of transient clinical signs and precise 

recording of time of death. 

GROSS NECROPSY: 

On all animals including those found dead or 

sacrificed in moribund condition. A full series of 

tissues and organs are taken and preserved. 
P.5 

PROTOCOL #: 798.1150 

Acute Inhalation Toxicity 

Rat/inhalation 

06/24/91 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision 

Vet. 

Senior tech. 

set-up 

compound prep. 

necropsy 

data record keeping 

exposure 

sampling & grav.anal. 

exposure sys. development 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

exposure sys. development 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (22 5% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

16 

12 

3 

( 3 ) 

6 

42 

( 5 ; 

( 2 ; 

( 3 ' 

( 6 , 

( 4 

( 8 t 

{ 1 4

 v 

46 

( 5 

( 14 

(

 9 

( 8 

( io 

17 

( 2 

( 1 

(

 2 

( 2 

( 4 

( 1 

\ *"* 

12 

8 

4 

154 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8. 01 

6.86 

12.36 

9.16 

10.58 

Total 

Dollars 

$552 

$167 

$98 

$141 

$415 

$368 

$117 

$148 

$73 

$42 

$2,123 

$4,776 

$6,899 
P.6 

PROTOCOL #: 798.1150 

Acute Inhalatio n Toxicit y 

Rat/inhalatio n 

06/24/91 

Other Direct Costs 

No. of 

Units 

8 

45 

4 

80 

14 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$5.00 

Total 

Dollars 

Overtime (animal tech.) 

Animals 

Feed 

Bedding 

Disposables 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G§A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% cf Total Cost) 

ESTIMATED FINAL COST: 

$32 

$383 

$90 

$16 

$70 

$212 

$803 

$7,702 

$1,694 

$9,396 

$1,879 

$11,275 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $11,280 $9,400 - $13,340 

TOTAL: $11,280 $9,400 - $13,340 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.7 

PROTOCOL #: 798.1150 06/24/91 

Acute Inhalation Toxicity 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 5 male and 

5 female rats. 

- Study duration is 2 weeks. 
P.8 

798.2450 INHALATION TOXICITY 

STUDY DESIGN: 

Group Dose Level Number of Animals 

1 Control 10 males, 10 females 

2 Low Dose io males, 10 females 

3 Mid Dose 10 males, 10 females 

4 High Dose 10 males, 10 females 

ADDITIONAL ANIMALS: 

If interim sacrifices are planned the number of 

animals in each group should be increased by the 

number of animals scheduled to be sacrificed during 

the course of the study. 

A satellite group of 10 animals per sex may be treated 

with the high dose level for 90 days and observed for 

reversibility, persistence or delayed occurrence of 

toxic effects for a period of not less than 28 days. 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats. 

AGE OF ANIMALS: 

Young adults. 

ROUTE OF EXPOSURE: 

Inhalation. 

DURATION AND FREQUENCY OF EXPOSURE: 

Six hours per day, seven days per week for a period of 

90 days. 

BODY WEIGHT: 

All animals are weighed at initiation of the test, 

weekly thereafter and at death or sacrifice. 

FOOD CONSUMPTION: 

Measured weekly if abnormal body weight changes are 

observed. 

OBSERVATIONS AND PHYSICAL EXAMINATIONS: 

At least once daily. 

HEMATOLOGY: 

Five animals per sex per group prior to initiation of 

the test, after 30 days of exposure and just prior to 

termination. Evaluation includes hematocrit, 

hemoglobin concentration, total red blood cell count, 

total and differential white blood cell count and a 

measurement of clotting potential. 
P.9 

798.2450 INHALATION TOXICTY (CONT.) 

CLINICAL BIOCHEMISTRY: 

Five animals per sex per group prior to initiation of 

the test, after 30 days of exposure and just prior to 

termination. The parameters to be measured include 

tests for electrolyte balance, carbohydrate metabolism 

and liver and kidney function. 

URINALYSIS: 

Not required routinely. 

OPHTHALMOLOGICAL EVALUATION: 

Five animals per sex per group just prior to initiation 

and just prior to termination of the test. 

GROSS NECROPSY: 

On all animals including those found dead or 

sacrificed in moribund condition. A full series of 

tissues and organs are taken and preserved. 

ORGAN WEIGHTS: 

At the time of terminal necropsy, the liver, kidneys, 

adrenals, brain and gonads are weighed from all 

animals. 

HISTOPATHOLOGY: 

Complete histopathology is performed on the 

respiratory tract and on all organs and tissues 

taken at necropsy from all animals in the control and 

high dose groups. All gross lesions and suspected 

target organs are evaluated from all animals. 
P.10 

PROTOCOL #: 798.2450 07/19/90 

Subchronic Exposure 

Rat/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Bd. Cert. Pathologist 

slide evaluation ( 

necropsy supervision ( 

Q.A. Inspector 

Vet. 

Clinical Chemist 

Clinical lab. tech. 

Histologist 

Histology tech. 

Senior tech. 

set-up i 

compound prep. i 

necropsy 

data record keeping 

exposure 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

bleeding 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Hematologist; 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

NO. of 

Hours 

60 

61 

54 ) 

7 ) 

148 

12 

50 

30 

154 

80 

237 

10 ] 

15 ] 

7 

23 

182 

803 

10 

728 

40 

( 15 

[ 1° 

200 

[ 5 

( 1° 

( 15 

( 8 

( 15 

( io 

( 137 

60 

8 

41 

12 

15 

1971 

Hourly 

Wage 

34.53 

39.66 

13.94 

23.56 

10.72 

9.94 

10.40 

8.83 

9.87 

8.01 

6.86 

10.72 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$2,072 

$2,419 

$2,063 

$283 

$536 

$298 

$1,602 

$706 

$2,339 

$6,432 

$1,372 

$643 

$185 

$507 

$110 

$159 

$21,726 

$48,882 

$70,608 
P.11 

PROTOCOL #: 798.2450 07/19/90 

Subchronic Exposure 

Rat/inhalation 

Other Direct Costs 

No. of 

Units 

Unit 

Cost 

Total 

Dollars 

Overtime (animal tech.) 150.0 $4.01 $601 

Animals 100.0 $8.50 $850 

Feed 31.0 $22.50 $698 

Bedding 600.0 $0.20 $120 

Disposables 91.0 $5.00 $455 

Histology supplies 1.0 $1,440.00 $1,440 

Clinical path, supplies 1.0 $1,080.00 $1,080 

Vet. ophthalmologist 2.0 $1,000.00 $2,000 

Lab. supplies (10% of SubTot. D.L.) $2,173 

TOTAL OTHER DIRECT COSTS: $9,416 

TOTAL BEFORE G&A: $80,024 

G@A ( 22% of Total Cost) $17,605 

TOTAL BEFORE FEE: $97,629 

FEE ( 20.0% of Total Cost) $19,526 

ESTIMATED FINAL COST: $117,155 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $117,150 $97,790 - $138,470 

ANALYTICAL CHEM: $22,560 $6,080 - $39,040 

TOTAL: $139,710 $103,870 - $177,510 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.12 

PROTOCOL #: 798.2450 07/19/90 

Subchronic Exposure 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 10 male and 

10 female rats. 

- Study duration is 13 weeks. 
P.13 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Mouse/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision ( 

Vet. 

Senior tech. 

set-up ( 

compound prep. ( 

observations 1 

necropsy I 

data record keeping i 

exposure i 

c-section i 

fetal evaluation 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

breeding 

c-section 

fetal evaluation 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

32 

64 

7 

7 ) 

8 

213 

25 ) 

6 ) 

56 ) 

7 ) 

12 ) 

20 ) 

20 ) 

67 ' 

346 

25 

80 

86 

20 

28 

( 40 

( 67 

68 

( 13 

( 8 

( 12 

( 4 

( 8 

( 8 

( 15 

4 

78 

24 

8 

852 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$1,105 

$892 

$229 

$188 

$2,102 

$2,771 

$466 

$92 

$964 

$220 

$85 

$9,116 

$20,510 

$29,626 
P.14 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Mouse/inhalation 

Other Direct Costs 

Overtime (animal tech. ) 

Animals 

Feed 

Bedding 

Stains 

Fixatives 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

No. of 

Units 

32.0 

250.0 

11.0 

480.0 

1.0 

1.0 

42.0 

D.L. ) 

Unit 

Cost 

$4.01 

$2.80 

$22.50 

$0.20 

$100.00 

$600.00 

$5.00 

Total 

Dollars 

$128 

$700 

$248 

$96 

$100 

$600 

$210 

$912 

$2,993 

$32,619 

$7,176 

$39,795 

$7,959 

$47,754 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $47,750 $39,780 - $56,570 

ANALYTICAL CHEM: $850 $230 - $1,460 

TOTAL: $48,600 $40,010 - $58,030 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.15 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Mouse/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 20 female 

mice. 

- Study duration is 6 weeks. 
P.16 

SYNOPSIS OF PROTOCOL 

TYPE OF STUDY: Developmental Toxicity Study In Rats 

NUMBER OF ANIMALS: 100 Female Rats 

NUMBER OF GROUPS: 3 Treated And 1 Control Group (25 Females Per Group) 

METHOD OF COMPOUND ADMINISTRATION: Inhalation, 6 Hours Per Day 

DOSING PERIOD: Gestation Days 6 Through 15 

BODY WEIGHT/FEED CONSUMPTION: Weekly 

CESARIAN DELIVERY: Gestation Day 19 

FETAL EVALUATION: 50% Of Fetuses For Skeletal Evaluation; 50% Of Fetuses 

For Soft Tissue Evaluation 

GROSS NECROPSY: All Animals 

16 
P.17 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Rat/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision ( 

Vet. 

Senior tech. 

set-up ( 

compound prep. . i 

observations i 

necropsy i 

data record keeping 

exposure 

c-section 

fetal evaluation 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

breeding 

c-section 

fetal evaluation 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

32 

62 

7 

7 ) 

8 

203 

15 ) 

6 ) 

56 ) 

7 ] 

12 

20 

20 

67 

336 

15 

80 

86 

20 

( 28 

: 40 

( 67 

63 

( 8 

[ 8 

( 12 

( 4 

( 8 

( 8 

( 15 

4 

78 

24 

8 

825 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$1,105 

$864 

$229 

$188 

$2,004 

$2,691 

$432 

$92 

$964 

$220 

$85 

$8,875 

$19,968 

$28,842 
P.18 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Rat/inhalation 

Other Direct Costs 

Overtime (animal tech.) 

Animals 

Feed 

Bedding 

Stains 

Fixatives 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

No. of 

Units 

32.0 

150.0 

25.0 

480.0 

1.0 

1.0 

42.0 

D.L. ) 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$100.00 

$600.00 

$5.00 

Total 

Dollars 

$128 

$1,275 

$563 

$96 

$100 

$600 

$210 

$887 

$3,859 

$32,702 

$7,194 

$39,896 

$7,979 

$47,875 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $47,880 $39,960 - $56,580 

ANALYTICAL CHEM: $850 $230 - $1,460 

TOTAL: $48,730 $40,190 - $58,040 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.19 

PROTOCOL #: 798.4350 07/19/90 

Inhalation Developmental Toxicity 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 20 female 

rats. 

- Studv duration is 6 weeks. 
P.20 

798.3320 COMBINED CHRONIC TOXICITY/ ONCOGENICITY 

STUDY DESIGN: 

Group 

1 

2 

3 

4 

Dose Level 

Control 

Low Dose 

Mid Dose 

High Dose 

Number c 

Oncogenicity 

50 per sex 

50 per sex 

50 per sex 

50 per sex 

•)f Animals 

Chronic Toxj 

20 per 

20 per 

20 per 

20 per 

Lcit; 

sex 

sex 

sex 

sex 

ADDITIONAL ANIMALS: 

The animals in the chronic toxicity groups are 

scheduled for sacrifice after 12 months of treatment. 

If additional interim sacrifices are planned, the 

number of animals in each group should be increased by 

the number of animals scheduled to be sacrificed 

during the course of the study. 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats or mice. 

AGE OF ANIMALS: 

Approximately 42 days at the start of the study. 

DURATION AND FREQUENCY OF EXPOSURE: 

Seven days per week for at least 24 months for rats 

and 18 months for mice. 

ROUTE OF EXPOSURE: 

Specified by test rule, may be oral, dermal or 

inhalation. 

BODY WEIGHT: 

All animals; weekly for the first 13 weeks and every 4 

weeks thereafter. 

FOOD CONSUMPTION: 

All animals; weekly for the first 13 weeks and every 4 

weeks thereafter in tests where the test chemical is 

administered in the feed. Water consumption is 

measured under the same schedule when the test 

chemical is administered in the drinking water. 

OBSERVATIONS AND PHYSICAL EXAMINATIONS: 

Once daily with special emphasis on the appearance and 

progression of palpable tumors. 
P.21 

798.3320 COMBINED CHRONIC TOXICITY/ONCOGENICITY (CONT.) 

HEMATOLOGY: 

Twenty animals per sex per group after 3, 6, 12 and 18 

months of exposure and just prior to study 

termination. Evaluation includes hematocrit, 

hemoglobin concentration, total red blood cell, white 

blood cell and platelet counts. Differential white 

blood cell counts will be performed on all control and 

high dose animals at each interval. 

CLINICAL BIOCHEMISTRY: 

Ten animals per sex per group prior to initiation of 

the test, at a midpoint interval and just prior to 

termination. The parameters to be measured include 

tests for electrolyte balance, carbohydrate metabolism 

and liver and kidney function. 

URINALYSIS: 

Ten animal per sex per group at the same intervals as 

the hematology evaluations. Evaluations to include 

appearance, volume, specific gravity , protein, 

glucose, ketones, bilirubin, occult blood and 

microscopic evaluation of the sediment. 

OPHTHALMOLOGICAL EVALUATION: 

Ten animals per sex per group just prior to initiation 

and just prior to termination of the test. 

GROSS NECROPSY: 

On all animals including those found dead or 

sacrificed in moribund condition. A full series of 

tissues and organs are taken and preserved. 

ORGAN WEIGHTS: 

At the time of terminal necropsy, the liver, kidneys, 

adrenals, brain and gonads are weighed from at least 

10 animals per sex per group. 

HISTOPATHOLOGY: 

Complete histopathology is performed on all organs and 

tissues taken at necropsy from all animals in the 

control and high dose group as well as from all found 

dead and moribund sacrificed animals. Additionally, 

histopathological evaluation is performed on all gross 

legions, target organs and the lungs, liver and 

kidneys from all animals. 
P.22 

PROTOCOL #: 798.3320 

Combined Chronic Toxicity/Oncogenicity 

Rat/inhalation 

08/15/90 

Direct Labor 

Study Director-toxicologist(PhD) 

Bd. Cert. Pathologist 

slide evaluation ( 

necropsy supervision i 

Q.A. Inspector 

Vet. 

Histologist 

Histology tech. 

Senior tech. 

set-up 

compound prep. 

necropsy 

data record keeping 

exposure 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

bleeding 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Hematologist 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. Of 

Hours 

424 

458 

411 ) 

47 ) 

1539 

57 

1075 

560 

3312 

60 

122 

47 , 

159 

2924 

9772 

[ 60 

' 8772 

812 

105 

23 

2770 

30 

424 

636 

53 

106 

[ 424 

' 1097 

93 

12 

250 

90 

106 

20518 

Hourly 

Wage 

34.53 

39.66 

13.94 

23.56 

10.40 

8.83 

9.87 

8.01 

6.86 

10.72 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$14,641 

$18,164 

$21,454 

$1,343 

$11,180 

$4,945 

$32,689 

$78,274 

$19,002 

$997 

$277 

$3,090 

$824 

$1,121 

$208,002 

$468,003 

$676,005 
P.23 -

PROTOCOL #: 798.3320 

Combined Chronic Toxicity/Oncogenicity 

Rat/inhalation 

No. of Unit 

Other Direct Costs Units Cost 

Overtime (animal tech.) 1410 $4.01 

Animals 600 $8.50 

Feed 1273 $22.50 

Bedding 25440 $0.20 

Disposables 731 $5.00 

Histology supplies 1 $10,080.00 

Vet. ophthalmologist 3 $1,000.00 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G§A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE PRICE 

PROTOCOL ESTIMATE: $1,109,740 $925,590 

ANALYTICAL CHEM: $157,920 $42,560 

TOTAL: $1,267,660 $968,150 

08/15/90 

Total 

Dollars 

$5,647 

$5,100 

$28,643 

$5,088 

$3,655 

$10,080 

$3,000 

$20,800 

$82,013 

$758,018 

$166,764 

$924,782 

$184,956 

$1,109,738 

RANGE 

-$1,312,670 

- $273,280 

-$1,585,950 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.24 

PROTOCOL #: 798.3320 08/15/90 

Combined Chronic Toxicity/Oncogenicity 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 50 male and 

50 female rats as well as 3 dosed and 1 control group of 20 

male and 20 female rats in a satellite group. 

- Study duration is 104 weeks. 
P.25 

PROTOCOL #: 798.4420 

Developmental Toxicity (Preliminary 

Mouse/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision ( 

Vet. 

Senior tech. 

set-up ( 

compound prep. ( 

necropsy ( 

data record keeping i 

exposure ( 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

breeding 

litter bw./obs. 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

ComDuter coder 

SUBTOTAL DIRECT LABOR: 

Overhead.(225% of D.L.) 

TOTAL DIRECT LABOR 

Screen) 

No. of 

Hours 

26 

28 

5 

5 ) 

7 

47 

20 ) 

2 ) 

5 ) 

10 ] 

10 

149 

20 

40 

16 

15 

28 

( 30 

43 

( io 

( 4 

(

 7 

( 3 

( 7 

( 4 

( 8 

8 

32 

18 

7 

370 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

07/19/90 

Total 

Dollars 

$898 

$390 

$163 

$165 

$464 

$1,193 

$295 

$185 

$396 

$165 

$74 

$4,388 

$9,873 

$14,261 
P.26 

PROTOCOL #: 798.4420 

Developmental Toxicity (Preliminary Screen) 

Mouse/inhalation 

NO. Of 

Other Direct Costs Units 

Overtime (animal tech.) 13.0 

Animals 200.0 

Feed 7.0 

Bedding 260.0 

Disposables 32.0 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE 

PROTOCOL ESTIMATE: $22,960 

ANALYTICAL CHEM: $850 

TOTAL: $23,810 

Protocol Estimates are made by variances in 1 

overhead, G&A, and fee rates assessed within 

laboratories. 

Unit 

Cost 

$4.01 

$2.80 

$22.50 

$0.20 

$5.00 

PRICE 

$19,120 

$230 

$19,350 

the fringe 

the United 

07/19/90 

Total 

Dollars 

$52 

$560 

$158 

$52 

$160 

$439 

$1,420 

$15,681 

$3,450 

$19,131 

$3,826 

$22,957 

RANGE 

$27,200 

$1,460 

$28,660 

benefits, 

1 States 
P.27 

PROTOCOL #: 798.4420 07/19/90 

Developmental Toxicity (Preliminary Screen) 

Mouse/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 1 dosed and 1 control group of 30 female 

mice. 

- Study duration is 4 1/2 weeks. 
P.28 

PROTOCOL #: 798.4420 07/19/90 

Developmental Toxicity (Preliminary Screen) 

Rat/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision ( 

Vet. 

Senior tech. 

set-up ( 

compound prep. ( 

necropsy ( 

data record keeping i 

exposure ( 

Tech. 

set-up ( 

exposure 

body wt./obs. 

necropsy 

breeding 

litter bw./obs. 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

26 

26 

5 

5 ) 

' 7 

38 

H ) 

2 ) 

5 ) 

10 ) 

10 ) 

133 

11 

40 

16 

15 

21 

30 

47 

( 5 

( 7 

( io 

( 3 

( 7 

( 7 

( 8 

8 

32 

18 

7 

347 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$898 

$362 

$163 

$165 

$375 

$1,065 

$322 

$185 

$396 

$165 

$74 

$4,171 

$9,384 

$13,554 
PROTOCOL #: 798.4420 

Developmental Toxicity (Preliminary Screen) 

Rat/inhalation 

NO. Of 

Other Direct Costs Units 

Overtime (animal tech.) 13.0 

Animals 108.0 

Feed 15.0 

Bedding 390.0 

Disposables 32.0 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE 

PROTOCOL ESTIMATE: $22,720 

ANALYTICAL CHEM: $850 

' TOTAL: $23,570 

Protocol Estimates are made by variances in 1 

overhead, G&A, and fee rates assessed within 

laboratories. 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$5.00 

PRICE 

$18,980 

$230 

$19,210 

the fringe 

the United 

P.29 

07/19/90 

Total 

Dollars 

$52 

$918 

$338 

$78 

$160 

$417 

$1,963 

$15,517 

$3,414 

$18,930 

$3,786 

$22,717 

RANGE 

$26,830 

$1,460 

$28,290 

benefits, 

1 States 
P.30 

PROTOCOL #: 798.4420 07/19/90 

Developmental Toxicity (Preliminary Screen) 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 1 dosed and 1 control group of 30 female 

rats. 

- Study duration is 4 1/2 weeks. 
P.31 

SYNOPSIS OF PROTOCOL 

TYPE OF STUDY: Reproduction And Fertility Effects 

NUMBER OF ANIMALS: 200 Total (100 Males, 100 Females) 

NUMBER OF GROUPS: 3 Treated And 1 Control (25 Males, 25 Females/Group) 

NUMBER OF GENERATIONS: Two 

NUMBER OF LITTERS PER GENERATION: One 

METHOD OF COMPOUND ADMINISTRATION: Inhalation 

DOSING REGIMEN: Seven Days Per Week Throughout Study; P Males And Females 

Dosed For 10 Weeks Prior To And Through Breeding, F. Parental 

Animals For 8 Weeks Prior To And Through Breeding; Parental 

Females Dosed Through Gestation And Lactation 

IN-LIFE OBSERVATIONS: Twice Daily For Obvious Signs Of Toxicity; Physical 

Examinations Weekly 

BODY WEIGHT/FEED CONSUMPTION: Weekly For Parental Animals 

PUP BODY WEIGHTS: At Birth, And On Days 4, 7, 14, And 21^ Individual Pups 

LITTER CULLING: Day 4 To 4 Males And 4 Females Per Litter 

GROSS NECROPSY: All Parental Animals And All F_ Animals At Day 21 Of Lactation 

And F. Animals Not Selected As Parental 

TISSUE PRESERVATION: Reproductive And Target Organs On All Pj And F{ Animals 

Selected For Mating 

HISTOPATHOLOGY: Reproductive And Target Organs On All High Dose And Control 

P. And F. Animals Selected For Mating 

18 
P.32 

PROTOCOL #: 798.4700 07/19/90 

Reproduction and Fertility Effects 

Rat/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Bd. Cert. Pathologist 

slide evaluation ( 

necropsy supervision ( 

Q.A. Inspector 

Vet. 

Histologist 

Histology tech. 

Senior tech. 

set-up i 

compound prep. i 

observations ( 

necropsy 

data record keeping 

exposure 

Tech. 

set-up 

exposure 

body wt./obs . 

necropsy 

breeding 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

NO. Of 

Hours 

148 

240 

211 ) 

29 ) 

495 

23 

338 

176 

1414 

36 ) 

35 ) 

760 ; 

29 t 

56 

498 

2995 

36 

1992 

881 

44 

( 42 

620 

( 18 

( 49 

( 74 

( I9 

( 37 

( 49 

( 374 

8 

78 

24 

37 

6596 

Hourly 

Wage 

34.53 

39.66 

13.94 

23.56 

10.40 

8.83 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$5,110 

$9,518 

$6,900 

$542 

$3,515 

$1,554 

$13,956 

$23,990 

$4,253 

$185 

$964 

$220 

$391 

$71,100 

$159,974 

$231,074 
P.33 

PROTOCOL #: 798.4700 

Reproduction and Fertility Effects 

Rat/inhalation 

No. of 

Other Direct Costs Units 

Overtime (animal tech.) 148.0 

Animals 180.0 

Feed 134.0 

Bedding 2960.0 

Stains 1.0 

Fixatives 1.0 

Disposables 259.0 

Histology supplies 1. 0 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE 

PROTOCOL ESTIMATE: $369,430 

';- ANALYTICAL CHEM: $70,220 

TOTAL: $439,650 

Protocol Estimates are made by variances in 1 

overhead, G&A, and fee rates assessed within 

laboratories. 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$200.00 

$600.00 

$5.00 

$6,336.00 

PRICE 

$307,520 

$18,920 

$326,440 

the fringe 

the United 

07/19/90 

Total 

Dollars 

$593 

$1,530 

$3,015 

$592 

$200 

$600 

$1,295 

$6,336 

$7,110 

$21,271 

$252,345 

$55,516 

$307,860 

$61,572 

$369,432 

RANGE 

$437,880 

$121,510 

$559,390 

benefits, 

1 States 
P.34 

PROTOCOL #: 798.4700 07/19/90 

Reproduction and Fertility Effects 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 20 male and 

24 female rats. 

- Study duration is 35 weeks. 
P.35 

798.5265 SALMONELLA TYPHIMURIUM REVERSE MUTATION ASSAY 

STUDY DESIGN PER STRAIN: 

Group 

1 

2 

3 

4 

5 

6 

7 

Dose Level 

Vehicle Control 

Positive Control 

Concentration 1 

Concentration 2 

Concentration 3 

Concentration 4 

Concentration 5 

Conditions 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

NUMBER OF BACTERIAL STRAINS: 

Salmonella typhimurium strains TA 1535, TA 1537, TA 98 

and TA 100. 

POSITIVE CONTROLS: 

The positive control should be strain specific. 

NUMBER OF PLATES PER CONCENTRATION: 

Plating should be done in triplicate. 

CONDITIONS: 

With and without metabolic activation. Activation is 

achieved by the addition of the S-9 fraction from the 

liver of rodents treated with a metabolic activator 

such as Arochlor 1254. 

EVALUATION: 

Individual plates are counted for the number of 

revertant colonies. 
P.36 

PROTOCOL #: 798.5265 

The Salmonella Typhimurium Reverse Mutation Assay 

Direct plate incorporation method 

08/15/90 

Direct Labor 

No. of 

Hours 

5 

3 

8 

8 ) 

16 

6 ) 

10 ) 

3 

1 

36 

Hourly 

Wage 

26.44 

13.94 

17.30 

9.94 

9.16 

10.58 

Total 

Dollars 

$132 

$42 

$138 

$159 

$27 

$11 

$510 

$1,146 

$1,656 

Study Director-geneticist/microbio. 

Q.A. Inspector 

Geneticist 

plate counting 

Laboratory technician 

set-up 

agar overlay/plating 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 
P.37 

PROTOCOL #.: 798.5265 

The Salmonella Typhimurium Reverse Mutation Assay 

Direct plate incorporation method 

08/15/90 

Other Direct Costs 

No. Of 

Units 

1.0 

1.0 

2.0 

Unit 

Cost 

$25.00 

$300.00 

$5.00 

Total 

Dollars 

$25 

$300 

$10 

$51 

$386 

$2,042 

$449 

$2,491 

$498 

$2,989 

Bacteria 

Plates/media supplies 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

D.L. 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $2,990 $2,510 - $3,510 

ANALYTICAL CHEM: $850 $230 - $1,460 

TOTAL: $3,840 $2,740 - $4,970 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.38 

PROTOCOL #: 798.5265 08/15/90 

The Salmonella Typhimurium Reverse Mutation Assay 

Direct plate incorporation method 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Based on 5 strains, 5 cone, 2 positive and 1 negative 

control groups with confirming assay. 
P.39 

Group 

1 

2 

3 

4 

5 

6 

Dose Level 

Vehicle Control 

Positive Control 

Concentration 1 

Concentration 2 

Concentration 3 

Concentration 4 

798.5300 DETECTION OF GENE MUTATIONS IN SOMATIC 

CELLS IN CULTURE 

STUDY DESIGN PER STRAIN: 

Conditions 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

CELL LINES: 

A variety of cell lines are used in this assay 

including subclones of the Chinese hamster ovary 

cells, the L5178Y (mouse lymphoma) cells and V-79 

cells. 

NUMBER OF CULTURES PER CONCENTRATION: 

At least duplicate cultures should be run for each 

concentration and condition. 

CONDITIONS: 

With and without metabolic activation. Activation is 

• achieved by the addition a chemical known to stimulate 

the metabolic activity of the test system to the 

culture media. 

TEST PROCEDURES/EVALUATION: 

The cells are exposed to the test substance in 

culture media with and without metabolic activation 

for a period of 1 to 5 hours. At the end of the 

exposure period, the cells are washed and cultured to 

determine viability and the expression of mutated 

cells. The cells are then grown in selective media 

for the determination of the numbers of mutants and 

the efficiency of the cells for cloning. 
P.40 

PROTOCOL #: 798.5300 

Detection of Gene Mutations in Somatic Cells in Culture 

CHO/HGPRT cells 

08/15/90 

Direct Labor 

No. of 

Hours 

20 

8 

4 

8 

7 

48 

6 

16 

6 

20 

12 

6 

6 

2 

8 

97 

) 

) 

) 

) 

) 

) 

) 

) 

) 

Hourly 

Wage 

26.44 

13.94 

17.30 

9.94 

23.08 

9.16 

Total 

Dollars 

$529 

$98 

$830 

$119 

$46 

$73 

$1,696 

$3,815 

$5,510 

Study Director-geneticist/microbio. 

study supervision ( 

data evaluation ( 

report preparation ( 

Q.A. Inspector 

Geneticist 

set-up ( 

cell preparation ( 

exposure ( 

cell counts ( 

Laboratory technician 

glassware prep. ( 

media prep ( 

Biostatistician 

Report typing/processing 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 
k 

PROTOCOL #: 798.5300 

Detection of Gene Mutations in Somatic Cells in Culture 

CHO/HGPRT cells 

No. of Unit 

Other Direct Costs Units Cost 

CHO cells 1.0 $40.00 

Disposables 2.0 $5.00 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE PRICE 

PROTOCOL ESTIMATE: $8,390 $6,960 

ANALYTICAL CHEM: $850 $230 

TOTAL: $9,240 $7,190 

Protocol Estimates are made by variances in the fringe 

overhead, G&A, and fee rates assessed within the United 

laboratories. 

P.41 

08/15/90 

Total 

Dollars 

$40 

$10 

$170 

$220 

$5,730 

$1,261 

$6,991 

$1,398 

$8,389 

RANGE 

$9,980 

$1,460 

$11,440 

benefits, 

States 
P.42 

PROTOCOL #: 798.5300 08/15/90 

Detection of Gene Mutations in Somatic Cells in Culture 

CHO/HGPRT cells 

ASSUMPTIONS: 

- Does not include cost for analytical method development or 

validation. 

- Based on single cultures at each level, triplicate plates 

per dose for cytotoxicity. 

- Five plates per dose evaluated for mutation. 

- Five dose levels plus positive and negative controls. 

- All groups tested with and without metabolic activation. 
P.43 

Group 

1 

2 

3 

4 

5 

6 

Dose Level 

Vehicle Control 

Positive Control 

Concentration 1 

Concentration 2 

Concentration 3 

Concentration 4 

798.5375 IN VITRO MAMMALIAN CYTOGENETICS 

STUDY DESIGN PER STRAIN: 

Conditions 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

With/Without Activation 

CELL LINES: 

A variety of cell lines and primary cell cultures can 

be used in this assay. Chinese hamster ovary and 

human lymphocytes are frequently used. 

NUMBER OF CULTURES PER CONCENTRATION: 

At least two for each experimental harvest time point 

with four time points. 

CONDITIONS: 

With and without metabolic activation. Activation is 

achieved by the addition a chemical known to stimulate 

the metabolic activity of the test system to the 

culture media. 

EVALUATION: 

Prior to harvest, a mitotic spindle inhibitor is added 

to the cell culture to stop the cells in metaphase. 

Chromosomal preparations are made and the slides are 

evaluated. The different types of chromosomal 

aberrations are enumerated for the control and treated 

groups. 

I 
P.44 

PROTOCOL #: 798.537 5 08/15/9 0 

In Vitro Mammalian cytogenetics 

Chromosone Aberrations/CHO cells 

Direct Labor 

Study Director-geneticist/microbio 

study supervision 

data evaluation 

report preparation 

Q.A. Inspector 

Geneticist 

set-up 

cell preparation/maintenance 

staining 

scoring 

Laboratory technician 

set-up 

cell preparation/maintenance 

staining 

scoring 

Biostatistician 

Report typing/processing 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. Of 

Hours 

16 

( 4 

( 4 

( 8 

8 

36 

( 6 

( io 

( 6 

( 14 

34 

( 4 

( 20 

( 2 

( 8 

2 

4 

100 

) 

) 

) 

) 

) 

) 

) 

) 

) 

) 

) 

Hourly 

Wage 

26.44 

13.94 

17.30 

9.94 

23.08 

9.16 

Total 

Dollars 

$423 

$112 

$623 

$338 

$46 

$37 

$1,578 

$3,551 

$5,129 
PROTOCOL #: 798.5375 

In Vitro Mammalian Cytogenetics 

Chromosone Aberrations/CHO cells 

No. of Unit 

Other Direct Costs Units Cost 

CHO cells 1.0 $40.00 

Disposables 2.0 $5.00 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE PRICE 

PROTOCOL ESTIMATE: $7,810 $6,480 

ANALYTICAL CHEM: $850 $2 3 0 

TOTAL: $8,660 $6,710 

Protocol Estimates are made by variances in the fringe 

overhead, G&A, and fee rates assessed within the United 

laboratories. 

P.45 

08/15/90 

Total 

Dollars 

$40 

$10 

$158 

$208 

$5,337 

$1,174 

$6,511 

$1,302 

$7,813 

RANGE 

$9,300 

$1,460 

$10,760 

benefits, 

States 
P.46 

PROTOCOL #: 798.5375 

In Vitro Mammalian Cytogenetics 

Chromosone Aberrations/CHO cells 

08/15/90 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Based on two cultures at each level, 24 hr. screening. 

- Four dose levels plus positive and negative controls. 

- All groups tested with and without metabolic activation. 
P.47 

798.5385 IN VIVO MAMMALIAN BONE MARROW CYTOGENETICS TESTS: 

CHROMOSOMAL ANALYSIS 

STUDY DESIGN: 

Group 

1 

2 

3 

4 

5 

Dose Level 

Negative Control 

Positive Control 

Low Dose 

Mid Dose 

High Dose 

Number of Animals 

15 males, 15 females 

15 males, 15 females 

15 males, 15 females 

15 males, 15 females 

15 males, 15 females 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats, mice or hamsters 

AGE OF ANIMALS: 

Sexually mature, young adults. 

FREQUENCY OF EXPOSURE: 

A single exposure is most widely used but other 

treatment schedules may be used. 

ROUTE OF EXPOSURE: 

Oral, via gavage, or intraperitoneal injection. Other 

routes can be selected, if appropriate. 

SACRIFICE SCHEDULE: 

Five males and five females from each group are 

sacrificed a 6, 24 and 48 hours after dosing. 

EVALUATION: 

Bone marrow samples are taken from each animal and 

slides are prepared for evaluation. Chromosomal 

changes in the cells in metaphase are evaluated from 

each animal. 
P.48 

PROTOCOL #: 798.5385 07/20/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Chromosomal Analysis 

Rat/inhalation 

Direct Labor 

Study Director-geneticist/microbio. 

Q.A. Inspector 

Vet. 

Clinical lab. tech. 

Geneticist 

Senior tech. 

set-up ( 

compound prep. 

necropsy 

data record keeping 

exposure 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. Of 

Hours 

36 

28 

6 

75 

50 

35 

is ; 

3 ' 

13 

2 

2 

66 

15 

8 

5 

( 38 

29 

8 

(

 4 

( 6 

( 2 

( 3 

(

 4 

( 2 

2 

30 

12 

2 

371 

Hourly 

Wage 

26.44 

13.94 

23.56 

9.94 

17.30 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$952 

$390 

$141 

$746 

$865 

$345 

$529 

$199 

$46 

$371 

$110 

$21 

$4,715 

$10,609 

$15,324 
P.49 

PROTOCOL #: 798.5385 07/20/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Chromosomal Analysis 

Rat/inhalation 

Other Direct Costs 

No. of 

Units 

6.0 

180.0 

11.0 

240.0 

300.0 

2.0 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$2.00 

$5.00 

Total 

Dollars 

$24 

$1,530 

$248 

$48 

$600 

$10 

$472 

$2,931 

$18,255 

$4,016 

$22,271 

$4,454 

$26,726 

Overtime (animal tech.) 

Animals 

Feed 

Bedding 

Stains/slides 

Disposables 

Lab. supplies (10% of SubTot, 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

D.L. ) 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $26,730 $22,390 - $31,470 

TOTAL: $26,730 $22,390 - $31,470 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.50 

PROTOCOL #: 798.5385 07/20/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Chromosomal Analysis 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed, 1 positive and 1 negative 

control groups of 5 male and 5 female rats for three 

sampling intervals at 6, 24 and 4 8 hours after treatment. 
P.51 

PROTOCOL 798.5395 07/20/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Micronucleus Assay 

Mouse/inhalation 

Direct Labor 

Study Director-geneticist/microbio. 

Q.A. Inspector 

Vet. 

Clinical lab. tech. 

Geneticist 

Senior tech. 

set-up 

compound prep. 

necropsy 

data record keeping 

exposure 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, 

feeding 

chamber cleaning 

Biostatistician 

etc. 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% 

TOTAL DIRECT LABOR 

of D.L.) 

No. of 

Hours 

36 

27 

6 

75 

50 

35 

15 ) 

3 ) 

13 

2 ' 

2 

66 

15 

8 

5 

38 

22 

( 8 

( 2 

( 3 

( 2 

( 3 

( 2 

( 2 

2 

30 

12 

2 

363 

Hourly 

Wage 

26.44 

13.94 

23.56 

9 .94 

17.30 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$952 

$376 

$141 

$746 

$865 

$345 

$529 

$151 

$46 

$371 

$110 

$21 

$4,653 

$10,470 

$15,123 
P.52 

PROTOCOL #: 798.5395 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Micronucleus Assay 

Mouse/inhalation 

No. of 

Other Direct Costs Units 

Overtime (animal tech.) 6.0 

Animals 180.0 

Feed 3.0 

Bedding 120.0 

Stains/slides 300.0 

Disposables 2.0 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE 

PROTOCOL ESTIMATE: $24,620 

TOTAL: $24,620 

Protocol Estimates are made by variances in 1 

overhead, G&A, and fee rates assessed within 

laboratories. 

Unit 

Cost 

$4.01 

$2.80 

$22.50 

$0.20 

$2.00 

$5.00 

PRICE 

$20,520 

$20,520 

the fringe 

the United 

07/20/90 

Total 

Dollars 

$24 

$504 

$68 

$24 

$600 

$10 

$465 

$1,695 

$16,818 

$3,700 

$20,517 

$4,103 

$24,621 

RANGE 

$29,140 

$29,140 

benefits, 

1 States 
P.53 

PROTOCOL #: 798.5395 07/20/90 

In Vivo Mammalian Bone Marrow Cytogenetics Tests: 

Micronucleus Assay 

Mouse/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed, 1 positive and 1 negative 

control groups of 5 male and 5 female mice for three 

sampling intervals at 6, 24 and 48 hours after treatment. 
P.54 

PROTOCOL #: 798.5550 

Unscheduled DNA Synthesis in Mammalian Cells in Culture 

CELL TYPE: 

Rat Hepatocytes 

TEST CONCENTRATIONS: 

Five test article concentrations, positive 

and negative controls. 

RADIOTRACER DETERMINATION: 

Autoradiography. 

NUMBER OF CULTURES: 

Six per concentration. 

CELL COUNTING: 

Fifty (50) cells per culture. 
J&J55_ 

PROTOCOL #: 798.5550 

Unscheduled DNA Synthesis in Mammalian Cells in Culture 

Primary cultures of rodent hepatocytes 

06/24/91 

Direct Labor 

No. of 

Hours 

Hourly 

Wage 

Total 

Dollars 

Study Director-geneticist/raicrobio. 

study supervision 

data evaluation 

report preparation 

Q.A. Inspector 

Geneticist 

set-up 

cell preparation/maintenance 

staining 

scoring 

Laboratory technician 

set-up 

cell preparation/maintenance 

staining 

scoring 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

Biostatistician 

Report typing/processing 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

32 

16 

8 

8 

9 

36 

6 

10 

6 

14 

34 

4 

20 

8 

n l 

1 

1 

1 i 

<z 

1 

2 

4 

115 

26.44 

13.94 

17.30 

9.94 

6.86 

23.08 

9.16 

$846 

$125 

$623 

$338 

$48 

$46 

$37 

$2,063 

$4,642 

$6,705 
P.56 

PROTOCOL #: 798.5350 06/24/91 

Unscheduled DNA Syntnesis in Mammalian Cells in Culture 

Primary cultures of rodent hepatocytes 

Other Direct Costs 

Animals 

Feed 

Bedding 

Media & Stains 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

No. of 

Units 

2.0 

1.0 

40.0 

1.0 

2.0 

D.L.) 

unit 

Cost 

$8.50 

$22.50 

$0.20 

$200.00 

$5.00 

Total 

Dollars 

$17 

$23 

$8 

$200 

$10 

$206 

$464 

$7,169 

$1,577 

$8,746 

$1,749 

ESTIMATED FINAL COST: $10,495 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $10,500 $8,720 - $12,460 

TOTAL: $10,500 $8,720 - $12,460 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.57 

PROTOCOL #: 798.5550 06/24/91 

Unscheduled DNA Synthesis in Mammalian Cells in Culture 

Primary cultures of rodent hepatocytes 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical chemistry. 

- Based on six cultures at each level, 24 hr. screening. 

- Five dose levels plus positive and negative controls. 

- All groups tested without metabolic activation. 

- UDS is measured by autoradiography technigue. 
P.58 

PROTOCOL #t 798.5900 

In Vitro Siste r Chromatid Exchange Assay 

CELL TYPE: 

Chinese Hamster Ovary 

TEST CONCENTRATIONS: 

Five test article concentrations, positive 

and negative controls. 

SCE DETERMINATION: 

Fluorescence plus Giemsa Method 

NUMBER OF CULTURES: 

At least two independent cultures per data point. 

CELL EXPOSURE: 

Cells should be exposed to the test chemical during at 

least two time intervals, e.g., GO and S phases. 
P.59 

PROTOCOL #: 798.5900 06/24/91 

In Vitro Sister Chromatid Exchange Assay 

Chinese Hamster Ovary Cells 

Direct Labor 

Study Director-geneticist/microbio. 

study supervision 

data evaluation 

report preparation 

Q.A. Inspector 

Geneticist 

set-up 

cell preparation/maintenance 

staining 

scoring 

Laboratory technician 

set-up 

cell preparation/maintenance 

staining 

scoring 

Biostatistician 

Report typing/processing 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

34 

( 16 

( 6 

( 12 

9 

36 

( 6 

( 12 

( 6 

( 12 

40 

( 6 

2 4 

( 2 

( 8 

2 

4 

116 

) 

) 

) 

) 

) 

) 

) 

) 

) 

) 

) 

Hourly 

Wage 

26.44 

13.94 

17.30 

9.94 

23.08 

9.16 

Total 

Dollars 

$899 

$125 

$623 

$398 

$46 

$37 

$2,128 

$4,787 

$6,915 
P.60 

PROTOCOL #: 798.5900 

In Vitro Sister Chromatid Exchange Assay 

Chinese Hamster Ovary Cells 

06/24/91 

Other Direct Costs 

No. of 

Units 

Unit 

Cost 

Total 

Dollars 

$200 

$10 

$213 

$423 

$7,338 

$1,614 

$8,952 

$1,790 

$10,742 

Media & stains 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% Of Total Cost) 

ESTIMATED FINAL COST: 

1.0 

2.0 

D.L. ; 

$200.00 

$5.00 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $10,740 $8,920 - $12,760 

TOTAL: $10,740 $8,920 - $12,760 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.61 

PROTOCOL #: 798.5900 

In Vitro Sister Chromatid Exchange Assay 

Chinese Hamster Ovary Cells 

06/24/91 

ASSUMPTIONS: 

- Does not include cost 

- Does not include cost 

- Based on two cultures 

- Five dose levels plus 

for dose range finding studies, 

for analytical chemistry, 

for each experimental point, 

positive and negative controls. 

- All groups tested with and v/ithout metabolic activation. 
PROTOCOL #: 798.5915 

In Vivo Sister Chromatid Exchange Assay 

STUDY DESIGN: 

Group 

l 

2 

3 

Dose Level 

Low Dose 

Mid Dose 

High Dose 

06/25/91 

vssay 

Number of Animals 

5 males, 5 females 

5 males, 5 females 

5 males, 5 females 

P.62 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats. 

AGE OF ANIMALS: 

Young adult animals. 

DURATION AND FREQUENCY OF EXPOSURE: 

A single exposure over a period of at least 4 hours 

followed by BdrU injections and treatment with a spindle 

inhibitor 2 hours prior to sacrifice. 

ROUTE OF EXPOSURE: 

Inhalation, for at least 4 hours. 

CELL TYPE: 

Bone marrow 

TEST CONCENTRATIONS: 

Three test article concentrations, positive 

and negative controls. 
P.63 

PROTOCOL 0i 798.5915 

In Vivo Sister Chromatid Exchange Assay 

Rat/inhalation 

06/25/91 

Direct Labor 

Study Director-geneticist/microbio, 

Q.A. Inspector 

Vet. 

Clinical lab. tech. 

Geneticist 

Senior tech. 

set-up 

compound prep. 

necropsy 

data record keeping 

exposure 

Tech'. 

set-up 

exposure 

body wt./obs. 

necropsy 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT LABOR 

No. of 

Hours 

36 

15 

6 

25 

17 

18 

( 5 ) 

( 3 ) 

(

 4 ) 

( 2 

( 4 ) 

34 

( 5 ] 

( 14 

( 2 

( 13 ' 

20 

( 3 

( 2 

( 3 

( 2 

( 3 

( 2 

( 5 

2 

30 

12 

1 

202 

Hourly 

Wage 

26.44 

13.94 

23.56 

9.94 

17.30 

9.87 

8.01 

6.86 

23.08 

12.36 

9.16 

10.58 

Total 

Dollars 

$952 

$209 

$141 

$249 

$294 

$178 

$272 

$137 

$46 

$371 

$110 

$21 

$2,980 

$6,705 

$9,685 
P.64 

PROTOCOL #: 798.5915 

In Vivo Sister Chromatid Exchange Assay 

Rat/inhalation 

06/25/91 

Other Direct Costs 

Overtime (animal tech.) 

Animals 

Feed 

Bedding 

Stains/slides 

Disposables 

Lab. supplies (10% of SubTot. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

No. of 

Units 

6.0 

60.0 

4.0 

120.0 

100,0 

2.0 

D.L.) 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$2.00 

$5.00 

Total 

Dollars 

$24 

$510 

$90 

$24 

$200 

$10 

$298 

$1,156 

$10,842 

$2,385 

$13,227 

$2,645 

$15,872 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $15,870 $13,240 - $18,780 

TOTAL: $15,870 $13,240 - $18,780 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United states 

laboratories. 
P.65 

PROTOCOL #: 798.5915 06/25/91 

In Vivo Sister Chromatid Exchange Assay 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical chemistry. 

- Study composed of 3 dosed, 1 positive and l negative 

control groups of 5 male and 5 female rats. 

- Animals administered BrdU after exposure to test chemical and 

a spindle inhibitor 2 hours prior to sacrifice. 

- After sacrifice, bone marrow is extracted and slides made and 

prepared for SCE evaluation. 
P.66 

PROTOCOL #: 798.6050 07/19/90 

Functional Observational Battery (Acute) 

Rat/inhalation 

Direct Labor 

Study Director-toxicologist(PhD) 

Q.A. Inspector 

Vet. Pathologist 

necropsy supervision ( 

Vet. 

Senior tech. 

set-up i 

compound prep. I 

necropsy i 

data record keeping i 

exposure i 

FOB testing i 

Tech. 

set-up 

exposure 

body wt./obs. 

necropsy 

FOB testing 

Animal care 

set-up 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

Overhead (225% of D.L.) 

TOTAL DIRECT,LABOR 

NO. Of 

Hours 

16 

18 

7 

7 ) 

6 

42 

9 ) 

2 ) 

7

 ; 

6 ) 

2 ) 

16 ] 

98 

9

 1 

8 

18 

I 15 

( 48 

23 

[ 5 

( 3 

4 

( 2 

( 4 

( 3 

:

 2 

21 

8 

4 

243 

Hourly 

Wage 

34.53 

13.94 

32.69 

23.56 

9.87 

8.01 

6.86 

12.36 

9.16 

10.58 

Total 

Dollars 

$552 

$251 

$229 

$141 

$415 

$785 

$158 

$260 

$73 

$42 

$2,906 

$6,539 

$9,445 

•T-.KS>--v--
PROTOCOL #: 798.6050 

Functional Observational Battery (Acute) 

Rat/inhalation 

No. of 

Other Direct Costs Units 

Overtime (animal tech.) 8 

Animals 90 

Feed 7 

Bedding 160 

Disposables 14 

Lab. supplies (10% of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE 

PROTOCOL ESTIMATE: $15,800 

TOTAL: $15,800 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$5.00 

PRICE 

$13,200 -

$13,200 -

Protocol Estimates are made by variances, in the fringe 

overhead, G&A, and fee rates assessed within 

laboratories. 

•••'•.•v-'-A. •-' .•-£*>•"--.•-:..•--s-rf^ .^•-••;-»':<iA .-..-A'MifjijV.rViis-:-': 

P.67 

07/19/90 

Total 

Dollars 

$32 

$765 

$158 

$32 

$70 

$291 

$1,347 

$10,792 

$2,374 

$13,166 

$2,633 

$15,799 

RANGE 

$18,660 

$18,660 

benefits, 

the United State*-

r :•• - .:..- ~ . 

'^^izZyiyy* •?'•*. 

. . ^i- - ... . 

- «H* A •:*,.«*-. 
P.68 

PROTOCOL #: 798.6050 07/19/90 

Functional Observational Battery (Acute) 

Rat/inhalation 

ASSUMPTIONS: 

- Does not include cost for dose range finding studies. 

- Does not include cost for analytical method development or 

validation. 

- Study composed of 3 dosed and 1 control group of 10 male and 

10 female rats. 

- Study duration is 2 weeks. 
P.69 

798.6050 FUNCTIONAL OBSERVATIONAL BATTERY 

STUDY DESIGN: 

Group 

1 

2 

3 

4 

Dose Level 

Control 

Low dose 

Mid dose 

High dose 

Number of Animals 

10 males, 10 females 

10 males, 10 females 

10 males, 10 females 

10 males, 10 females 

SATELLITE GROUPS: 

A satellite group may be treated with the high dose 

level for the duration of exposure and observed for 

reversibility, persistence or delayed occurrence of 

toxic effects for a post-treatment period of 

appropriate duration, normally not less than 28 days 

(not included in the cost estimate). 

ANIMAL SPECIES OR STRAIN: 

Laboratory rats. 

AGE OF ANIMALS: 

At least 42 days 

DURATION AND FREQUENCY OF EXPOSURE: 

Daily dosing, seven days per week for 13 weeks. 

ROUTE OF EXPOSURE: 

Dermal, non-occluded. 

OTHER: 

Concurrent or historical data from the laboratory 

performing the test to provide evidence that the 

observation procedures detect neurotoxic changes. 

The tests may be performed on subjects from other 

toxicity tests provided that the requirements of 

neither study are violated by combination. 

BODY WEIGHTS: 

Weekly, throughout the study. 

OBSERVATION INTERVALS: 

Prior to study initiation, 1, 6 and 24 hours after 

initial treatment, days 7 and 14 of study and monthly 

thereafter. 

OBSERVATIONS: 

A. Any unusual responses with respect to body position, 

activity level, coordination and gait. 

B. Unusual or bizzare behavior such as headflicking, head 

searching, compulsive biting or licking, self 

mutilation, circling and walking backwards. 
P.70 

c. 

D. 

toiicitv ll'D? SUC h " th °

S e t3nPie«Uy observed in toxicity studies and considered related to 

slllvatfnn31 Chan ?

e s ' e ' 8 - increased lacrimation or 

const*!.??'

 con ^^i°n s or tremors, pupil dilation or 

diarrhl! ,'

 chan «e s i n respiration or urination, 

diarrhea, piloerection and vocalization. 

Forelimb and hindliab grip strength. 

E" ^uictiin

 SllX* °f te ;

t £ t 0 evaluat e •"•«*« on sensor 

i»ef„i ; r f

fl8Xe s an d Pa i n Perception. These may 

include tests such as auditory startle response 

Ticking

 refl6X ' ^ i n " y "«« . hot plaE"; * 

balancf MnS??8 !'

 b flanC B bea m •1«i"bri»«. rotorod 

r!.««n

 h fndllB b sPi*y measurements, visual placing 

response, toe and/or tail pinch response, visual 

<no-i?fuIMPO ?"# , '

n d reactio n t o Poaitional changes (positive geotropism and awkward positions). 

ASSUMPTIONS: 

hlstorirlf

 h " Hi l °f th e re «uir *d equipment and 

of thii"estrPerienC e a "

d COntro i dat a fo r th e conduct 

y 
P.71 

CFR 79B.6050 03 JUN 1990 

PRGTOCDL LSTIMATE 

FUNCTIONAL OBSERVATIONAL DATTERY 

Rat .' Inhalation 

Subcronic 

Direc t Labor 

Num. Of 

Hour s 

Hour 1y 

Waae Tota l Dollars 

Btuuv Dir Toxicoloqist(PhD ) 

.A r-t. ;,i=aectar 

ete n nar y Pathologis t 

Necrops y supervisio n 

_aa An i rna 1 Vet et" inar i an 

Senior i-irtirnal Technicia n 

Sec up 

Compound preparatio n 

Data recor d keeping 

Necrops v 

Exposure 

FOB testin g 

r-.i \ i inai Tecnn i c i an 

. OIJ -. .vrfi ul'its / Ob ser vat i oris 

. ) fi L r G i j 3 V 

:..Gu:ui' e 

•_ o _ .= •= C i ("iCj 

ni'lliiia l Jai'Stdlket " 

;":ocii, -set up 

Deddina change 

!"•>ooi"n iiiai n t e n ance 

Caue cleanin g 

Feedi ng 

hashing feeders, etc. 

ChaitiDer cleaning 

Diostati sti cian 

f-.eoort Writer/Data Coordinator 

Word Processor/Typist 

Computer Coder 

SUBTOTAL DIRECT LABOR: 

\ 

( 

( 

( 

( 

( 

( 

\ 

( 

I. 

(. 

( 

( 

( 

s 

i 

( 

( 

60. 00 

;31.00 

7. 00 

7.00) 

12.00 

262.00 

a. oo) 

15.00) 

23.00) 

7. 00) 

182.00) 

30.00) 

394.00. 

3.00) 

o3.00) 

15.00) 

72B.OO) 

i-30.00) 

199.00 

4. 00) 

10.00) 

3. 00) 

15.00) 

10.00) 

15.00) 

137.00) 

8. 00 

33.00 

20. 00 

15.00 

34.53 

13.94 

32.69 

23.56 

9.37 

Q. 01 

5.86 

23.03 

9. 16 

10.53 

,071.30 

,12?.14 

228.83 

2B2.72 

:.535.?4 

.", 160.94 

J. . JUwJ, 14 

184.64 

407.38 

183.20 

.158.70 

$15,758.93 

Overhead (225.0'/. of D.L. ) 

TOTAL DIRECT LABOR: $51,216.5 2 
P.72 

PROTOCOL #: 40 CFR 798.6050 08 JUN 1990 

FUNCTIONAL OBSERVATIONAL DATTERY 

Rat / Inhalation 

Subcronic 

Other Direct Costs 

Overt ijie: 

rimmal Technician 

Rats 

Feed 

Eiedding 

Disposables 

Lab Supplies ( 10.07. of SubTot. D.L.) 

TOTAL OTHER DIRECT COSTS: 

TOTAL COOT DEFORE GS<A: 

G&A ( 22.07. of Total Cost) 

"OTAL COOT BEFORE FEE: 

FEE ;' 20.07. of Total Cost) 

Num. Of 

Units 

150.00 

90.00 

24. 00 

600.00 

91. 00 

.L.) 

Unit 

Cost 

4.01 

8.50 

22.50 

0. 20 

5.00 

1575.89 1 

$4 

600.75 

765.00 

540.00 

120.00 

455.00 

,575.89 

.056.64 

ISTITIATED FINAL COST: 

-••• O O a -^. > •-> • X > 

12.160.10 

•-E67. 433 . 2 7 

13.486.65 

£80.919.92 

PROTOCOL ESTIMATE: 

ANALYITCAL CHEM.: 

TOTAL: 

COST COMPARISON 

BEST ESTIMATE 

$80,920 

$23,410 

$104,330 

PRICE RANGE 

$6: SOO 96,000 

$6,310 - 40,500 

$73,610 - 136,500 

;:,sjj"ft>*. 
P.73 

PROTOCOL its 40 CFR 798.6050 08 JUN 1790 

FUNCTIONAL OBSERVATIONAL BATTERY 

R at / Inhalatio n 

Subcronic 

Protocol Estimates are made by variances in the fringe benefit: 

overhead, G&A, and fee rates assesed within the United States 

1aborstarles. 

ASSUMPTIONS: 

- 1 control & 3 treated groups ot 10 males and 10 females each. 

- No analytical chemistry validation of stability or dosage verification. 

ANALYTICAL CHEMISTRY COSTS: 

Analvtical Chemistry costs are estimated to range 

from : £ 6,310 to $ 40,500. 
P.74 

798.7100 METABOLISM 

STUDY DESIGN: 

Group 

1 

2 

3 

4 

Dose Level 

Low Dose 

Low Dose 

Low Dose 

High Dose 

itoute 

IV xl * 

OR xl ** 

OR xl4 *** 

OR xl 

Number of Animals 

4 males, 4 females 

4 males, 4 females 

4 males, 4 females 

4 males, 4 females 

single intravenous dose 

single oral dose 

*** daily oral dose for 14 days 

* 

* * 

ANIMAL 8PECIES OR STRAIN: 

Laboratory rats (dogs sometimes used). 

AGE OF ANIMALS: 

Young adults. 

NATURE OF THE CHEMICAL: 

Analytically pure, radiolabelled test chemical should 

be used. 

DURATION OF TEST: 

At least 7 days after administration of the 

radiolabelled test chemical or until 95% recovery of 

the administered dose is excreted. 

ROUTE OF EXPOSURE: 

Oral dosing may be achieved via gavage or capsule 

(generally in dog studies). The intravenous dose 

group is used for comparison to the oral dose groups 

to determine of the rate and degree of absorption of 

the test chemical. 

DOSING REGIMEN: 

All animals in groups 1,2 and 4 receive a single dose 

of the radiolabelled test chemical. The group 3 

animals receive single oral doses of nonradiolabelled 

material daily for 14 consecutive days. Twenty-four 

hours after the last dose, the group 3 animals receive 

a single oral dose of the radiolabelled test chemical. 
P.75 

798.7100 METABOLISM (CONT.) 

SAMPLE COLLECTION/ANALYSIS: 

Quantities of radiolabel in the urine, feces and 

expired air are determined in samples collected at 

4,8,12,24 and 36 hours, 2,3,4,5,6 and 7 days post 

dosing with the radiolabelled test chemical. 

At sacrifice, the brain, fat, gonads, bone, heart, 

kidneys, liver, lungs, muscle, spleen, tissues showing 

gross pathology and carcass are retained and the 

quantity of radiolabel determined. 

Appropriate analytical procedures are followed to 

determine to the extent of metabolism of the test 

chemical in urine and feces samples from all groups. 

The identification of the major metabolites is not 

required but may be requested. 
PROTOCOL #: 798.7100 

Metabolism 

Rat/inhalation 

Direct Labor 

Study Director-anal. chem. 

Q.A. Inspector 

Chemist 

extractions ( 

GC/TLC, GC/MS ( 

data collection/analysis 

analytical monitoring 

Chemistry tech. 

set-up 

radiocarbon analysis 

homogenization/combustion 

TLC/extractions 

data reduction 

data entry 

Senior tech. 

set-up 

compound prep. 

data record keeping 

sample collection 

exposure 

Tech. 

set-up 

exposure 

animal room activities 

body wt./obs. 

sample collection 

Chem. lab. assistant 

homogenization/combustion 

glassware prep./maintenance 

radiocarbon analysis 

Animal care 

set-up metab. cages 

bedding change 

cage cleaning 

room maintenance 

washing feeders, etc. 

feeding 

chamber cleaning 

Biostatistician 

Report/data coordinator 

Report typing/processing 

Computer coder 

SUBTOTAL DIRECT LABOR: 

No. of 

Hours 

60 

243 

295 

48

 , 

192 

48 

7 

1219 

16 

500 

173 

128 

134 

268 

48 

8 

1 

8 

13 

18 

135 

10 

72 

15 

12 

26 

1105 

173 

32 

900 

43 

5 

2 

3 

3 

5 

2 

23 

8 

40 

40 

3 

3239 

Hourly 

Wage 

21.63 

13.94 

12.74 

9.38 

9.87 

8.01 

7.09 

6.86 

23.08 

12.36 

9.16 

10.58 

08/16/90 

Total 

Dollars 

$1,298 

$3,387 

$3,758 

$11,434 

$474 

$1,081 

$7,834 

$295 

$185 

$494 

$366 

$32 

$30,639 

P.76 
P.77 

PROTOCOL #: 798.7100 

Metabolism 

Rat/inhalation 

08/16/90 

Other Direct Costs 

No. Of 

Units 

10.0 

40.0 

5.0 

100.0 

1.0 

1.0 

22.0 

1.0 

Unit 

Cost 

$4.01 

$8.50 

$22.50 

$0.20 

$1,500.00 

$200.00 

$5.00 

$6,000.00 

Total 

Dollars 

$40 

$340 

$113 

$20 

$1,500 

$200 

$110 

$6,000 

$3,064 

$11,387 

$110,965 

$24,412 

$135,377 

$27,075 

$162,452 

Overtime (animal tech.) 

Animals 

Feed 

Bedding 

Solvents 

TLC plates 

Disposables 

Counting accessories/autoradiogra 

Lab. supplies (10% of SubTot. D.L. 

TOTAL OTHER DIRECT COSTS: 

TOTAL BEFORE G&A: 

G@A ( 22% of Total Cost) 

TOTAL BEFORE FEE: 

FEE ( 20.0% of Total Cost) 

ESTIMATED FINAL COST: 

COST COMPARISON 

BEST 

ESTIMATE PRICE RANGE 

PROTOCOL ESTIMATE: $162,450 

TOTAL: $162,450 

$135,430 -

$135,430 -

$192,250 

$192,250 

Protocol Estimates are made by variances in the fringe benefits, 

overhead, G&A, and fee rates assessed within the United States 

laboratories. 
P.7S_ 

PROTOCOL #: 798.7100 

Metabolism 

Rat/inhalation 

08/16/90 

ASSUMPTIONS: 

TLC for quantification utilizes pooled samples from same 

sexed animals from each group and at each interval. 

GC/MS metabolite identification assumed on spots from three 

intervals. 

Exposure via the 

as a comparative 

Does not include 

Does not include 

validation. 

Does not include 

or for synthesis 

gavage route with an intravenous group 

control 

dose range finding studies. 

analytical method development 

cost 

cost 

for 

for or 

cost for radiolabelled material($10,000-20,000) 

of metabolites standards. 
P.79 

I. COST ESTIMATION PROCEDURES 

A. OVERVIEW 

The methodology discussed below describes the steps taken 

in the preparation of cost estimates for chemical fate, environmental effects and health effects tests as required by the Office 

of Toxic Substances, U.S. Environmental Protection Agency. This 

methodology has been developed from contractor experience in 

serving the testing needs of various governmental agencies and 

industry. 

The principle components of the cost estimates are total 

direct labor (TDL), Overhead (OH), Other Direct Costs (ODC), 

General and Administrative Cost (G&A) and Fee. Each of these 

components are discussed in detail below. 

B. TOTAL DIRECT LABOR 

Upon receipt of a task assignment, the program manager 

reviews the scope of work to determine the requirements to be 

met. If the assignment requires the costing of a standard test 

guideline, a copy of that guideline is secured, the test 

procedures within the guideline are reviewed and an outline of 

the study protocol is prepared. Occasionally, a test rule 

requires the combination of testing procedures from several tests 

or evaluation of journal articles for cost considerations. In 

these cases, a study design/protocol is prepared to comply with 

current regulatory requirements and reviewed for technical and 

logistical feasibility. Any questions that arise are passed 
P.80 

through the EPA/RIB contact to the appropriate technical 

personnel within the Agency. Upon completion of the study 

design, the outline of the test, as indicated above, is prepared. 

The outline is an itemized breakdown of the study design 

into the various components required by the protocol, e.g., the 

number of animals, duration of the test, dosing regime, specialized tests, etc. The outline is further subdivided into task 

components according to technical areas, e.g. animal care, histology, analytical chemistry, etc. The required tasks are 

reviewed and the usual time required for completion is estimated. 

After completion of this process for all tasks contained in the 

outline, a labor category/task performance sheet is completed. 

This sheet lists various job categories required by the test, the 

total number of labor hours for that category and a breakdown of 

the hours into the various tasks performed. 

The total labor hours for each job category are then multiplied by the hourly rate assigned to that labor category to 

determine the total labor dollars. The total direct labor (TDL) 

is the sum of the labor dollars for each job category. The salary 

rates currently in use are estimates of industry averages 

obtained from testing laboratories, government contractors, 

placement services and technical job placement advertisements in 

November, 1985. 
P.81 

C. OVERHEAD 

The overhead rate is applied as a percentage of TDL. 

This percentage represents those costs incurred by a laboratory 

in the conduct of its business; that is facility operation, 

fringe benefits and indirect labor. 

The costs associated with the operation of a testing laboratory facility include factors such as mortgage or rent payments, 

utilities, depreciation and amortization of non-expendable equipment, insurance, property taxes, repairs and maintenance of the 

facility and equipment. Indirect support costs are also included 

in overhead. These costs include items such as fees and 

licenses, publication, postage, computer expenses, travel, office 

supplies and materials, recruitment and relocation of personnel. 

Indirect labor costs, i.e., those associated with personnel 

who are required for the functioning of a laboratory but are not 

directly allotted to the study, are also included in overhead. 

Indirect labor includes security, maintenance and supervisory 

personnel. 

Because of the confidential nature of such financial information, private laboratories, as a practice, do not include overhead rates in their bids. These rates are, however, supplied to 

the government by contractors in the bidding process. Overhead 

rates in current use are those accepted by testing laboratories 

in bidding government contracts as well as rates calculated by 

Mathtech from laboratory competitive bids for various programs. 
P.82 

D. OTHER DIRECT COSTS 

Other direct costs (ODC) consist of laboratory and 

expendable supplies, subcontracted services and overtime premium. 

1) Laboratory and Expendable Supplies 

These supplies are those items required specifically for 

the test. They include animals and associated 

products (feed, bedding, animal room supplies), 

chemicals and reagents, and disposable supplies 

for specialized tests (histology, analytical chemistry). 

The costs for equipment to conduct the tests are not 

included. It is assumed that a laboratory conducting a 

specific test would have the major equipment required 

for that test. The allocation of costs associated with 

major equipment are included in the overhead calculation discussed above. 

2) Subcontracted Services 

Since it is not cost effective for a laboratory to have 

all of the required equipment or specific expertise to 

conduct a broad variety of tests, some service may be 

subcontracted. As an example, ophthalmological 

examination may be required one or two times per year by 

a specific laboratory requiring the services of a 

consulting ophthalmologist. These fees are included as 

an ODC. 

3) Overtime Premium 

Certain employees may be required to perform functions 

on tests that require more than eight hours per day or 

40 hours per week. The functioning of an animal testing 

laboratory requires seven day per week coverage. 

Overtime is included as an ODC. 

E. GENERAL AND ADMINISTRATIVE COSTS 

These costs are the salaries of the support services 

required in the administrative function of the company or 

corporation rather than the laboratory. These services include 

the accounting, personnel, administrative, purchasing, payroll, 
PJ33_ 

legal and marketing departments. The G&A rates are less variable 

than those for overhead and an average of 15% covers a broad 

range of various businesses. 

F. FEE 

The fee or profit assessed by a testing laboratory can 

vary markedly. Factors involved in determining a fee rate 

include the available capacity of the laboratory, specific 

capability of the laboratory, duration of the test, the type of 

test and current market conditions. 

G. FINAL COST DETERMINATION 

The initial step in the preparation of the cost estimate 

is the completion of the labor hour estimates and determination 

of the other direct costs as required by the protocol for the 

study. With this information, the cost estimation proceeds as 

follows: 

Total hours x hourly rate 

Overhead % x TDL 

% G&A x Total 1 above = 

% Fee x Total 2 above = 

Total Direct Labor (TDL) 

+ Total Overhead (OH) 

+ Other Direct Costs 

+ 

+ 

Total 

Total 

Total 

Total 

1 

G&A 

2 

Fee 

Estimated costs Sum of components above 
P.84 

Depending on the duration of the test, salary adjustments 

and inflation factors may be included for the TDL and the ODC, 

respectively, based on current government standards and the 

consumers price index. 

An estimated cost range is then calculated by substituting 

the lowest and highest expected overhead rates into the general 

calculation. Overhead is used as the main variable because it is 

the most significant cost component in the overall test cost and 

can vary greatly from company to company and even within specific departments within a company. G&A rates are generally consistent from the businesses surveyed. No consideration for 

market conditions are made so a reasonable fee rate is maintained 

in the estimation. Under slow market conditions, this component 

would, in fact, decrease in the contract laboratory business in 

order to increase the competitiveness of the bid to potential 

sponsors. 

Industrial laboratories show higher overhead rates than 

those used for the above estimation procedures. The estimated 

test costs between the industrial and contract laboratories, 

however, would be similar. The higher overhead rates of the 

industrial laboratories would be balanced by the fee charged by 

the contract laboratories,i.e., the industrial laboratories do 

not charge themselves a fee for doing studies in house. 
P.85 

II. ANALYTICAL CHEMISTRY REQUIREMENTS 

An integral part of the conduct of an acceptable test is the 

assurance that the test organisms are exposed to the appropriate 

dose or concentration of the test chemical. The dose or concentration levels can be directly calculated (nominal concentration 

or dose) based on the mixing procedure or the generation procedures. As an example, the amount of test chemical (mg) added to 

a measured amount of dosing vehicle or water (ml) calculates to a 

specific concentration (mg/ml). Because of the chemical or physical nature of the test chemical, the calculated nominal concentration may be several fold different from the actual concentration to which the organism is exposed. The test chemical may 

also rapidly degrade under specific circumstances so that the 

concentration may be correct at the start of the test, but significantly lower a few hours to several days later. 

In order to assure that the correct exposure occurs during 

specific tests, certain of the Toxic Substances Control Act testing guidelines specify the analysis of dosing samples to determine actual concentrations of exposure at several intervals 

during the test period. These requirements are specifically 

found in the test guidelines for the environmental effects tests 

and for the health effects tests by the inhalation route. 

Although not directly a part of the individual TSCA testing 

guidelines for other health effects tests, the requirement for 

analytical verification of test chemical purity, stability and 

appropriate concentration in dosing solutions are mandated by the 

TSCA Good Laboratory Practice Standards (40 CFR Part 792, 
P.86 

November 29, 1983). Of course, analytical determinations are the 

key aspects of bioconcentration and metabolism studies. 

Analytical chemistry determinations are comprised of two 

distinct phases: method validation/ verification and sample 

analysis. Each phase is are discussed below. 

The method validation/ verification phase involves the setup 

and trial analysis of known standards to assure that the prescribed methods work for both the testing laboratory and the 

specific matrix to be analyzed for test chemical content. 

Because of variations in techniques and equipment from one laboratory to another, it is essential that the laboratory analyzing 

the test samples demonstrates consistency of results with the 

laboratory that developed the methods. Different procedures may 

be required in the preparation of samples based on the matrix in 

which the test chemical is mixed. Samples in water may be 

directly analyzed with minimal preparation; samples from test 

chemical/ diet admixtures may require extensive extraction and 

cleanup prior to analysis. 

Method validation/ verification must be completed prior to 

the conduct of all tests. Once a laboratory has completed the 

method validation associated with a particular chemical in a 

specific matrix, the method validation does not need to be 

repeated for subsequent tests of a related nature. Thus, a 
P.87 

laboratory conducting several aquatic tests with a specific test 

chemical would be required to validate the method once and apply 

this method to the samples from several similar tests. 

Sample analysis schedules are stated in the environmental 

effects guidelines as well as the inhalation health effect 

guidelines so that an accurate number of required samples can be 

determined for each test. For the remaining health effects 

tests, certain assumptions must be made in determining the 

frequency for sampling and analysis as well as stability testing 

and homogeneity of mixing evaluations in order to comply with the 

GLPs. These areas of the GLPs are currently under revision and 

the numbers of samples required for analysis are expected to 

change. 

The costs for method validation/ verification and for sample 

analysis vary due to the degrees of complexity required in the 

preparation of the test sample to an analyzable form, the type of 

equipment necessary, the specifications related to the chemical 

under analysis and the suitability of the method to automation. 

The number of samples analyzed at each interval also is important 

to the cost. The instrument setup time, as an example, is the 

same whether one sample or multiple samples are analyzed. 

Standards are also analyzed regardless of the number of samples 

analyzed at a particular interval. If samples from several 

intervals can be saved and analyzed at the same time, some cost 

efficiencies would be realized. 
P.88 

The method validation cost ranges for various analytical 

methods are as follows: 

Gas chromatography/ Liquid chromatography $2200 to 7000 

Ultraviolet/Visible Absorbance Spectroscopy $ 550 to 1350 

Atomic Absorption Spectrophotometry $275 to 550 

The analytical cost per sample for the above methods are 

estimated as follows: 

Gas Chromatography/ Liquid Chromatography $ 57 to 122 

Ultraviolet/Visible Absorbance Spectroscopy $ 25 to 42 

Atomic Absorption Spectrophotometry $ 19 to 30 

Expected analytical costs, where available, are listed for 

individual guidelines in Attachment I. These costs also appear 

in the detail cost breakdowns of the guidelines which are 

contained in Appendices 5 and 6 (Volume I) and Appendix 7 (Volume 

II). A breif description of the three analytical methods listed 

above is also included as Attachment II. 

10 
ATTACHMENT I 

ENVIRONMENTAL EFFECTS TESTS 

P.89 

797.1050 

797.1160 

797.1300 

797.1330 

797.1350 

797.1400 

797.1520 

Algal acute toxicity test 

GC/LC $1710 to $3660 

UV/VIS $ 750 to $1260 

AA $ 570 to $ 900 

Lemna acture toxicity test 

GC/LC $2052 to $4392 

UV/VIS $ 900 to $1512 

AA $ 684 to $1080 

Daphnid acute toxicicty test 

Static (ST) 

GC/LC $1026 to $2196 

UV/VIS $ 450 to $ 765 

AA $ 342 to $ 540 

Flowthrough (FT) 

GC/LC $2280 to $4880 

UV/VIS $1000 to $1680 

AA $ 760 to $1200 

Daphnid chronic toxicity test 

Flowthrough (FT) 

GC/LC $5928 to $12688 

UV/VIS $2600 to $4368 

AA $1976 to $3120 

Daphnid chronic toxicity test 

GC/LC $4674 to $10004 

UV/VIS $2050 to $3444 

AA $1558 to $2460 

Fish acute toxicity 

Static (ST) 

GC/LC $1026 to $2196 

UV/VIS $ 450 to $ 756 

AA $ 342 to $ 540 

Flowthrough (FT) 

GC/LC $2280 to $4880 

UV/VIS $1000 to $1680 

AA $ 760 to $1200 

Fish bioconcentration test 

GC/LC $11400 to $22400 

UV/VIS $ 5000 to $ 8400 

AA $ 3800 to $ 6000 

30 samples 

36 samples 

18 samples 

40 samples 

104 samples 

82 samples 

18 samples 

40 samples 

200 samples 

11 
P.90 

797.1600 

ATTACHMENT I (Continued) 

ENVIRONMENTAL EFFECTS TESTS 

Fish early life stage toxicity test 

Minnows 

GC/LC 

UV/VIS 

AA 

Trout 

GC/LC 

UV/VIS 

AA 

$4902 to $10492 

$2150 to $3612 

$1634 to $2580 

$8322 to $17812 

$3650 to 6132 

$2774 to 4380 

797.1800 

797.1830 

797.1930 

797.1950 

Oyster acute toxicity test 

GC/LC $1596 to $3416 

UV/VIS $ 700 to $1176 

AA $ 532 to $ 840 

Oyster bioconcentration test 

GC/LC $9120 to $19520 

UV/VIS $4000 to $6720 

AA $3040 to $4800 

Mysid shrimp acute toxicity test 

Static (ST) 

GC/LC $1026 to $2196 

UV/VIS $ 450 to $ 756 

AA $ 342 to $ 540 

Flowthrough (FT) 

GC/LC $3078 to $6588 

UV/VIS $1350 to $2268 

AA $1026 to $1620 

Mysid shrimp chronic toxicity test 

GC/LC $6156 to $13176 

UV/VIS $2700 to $4536 

AA $2052 to $3240 

86 samples 

146 samples 

28 samples 

160 samples 

18 samples 

54 samples 

108 samples 

797.2750 Seed germination/root elongation toxicity test 

12 samples 

GC/LC $684 to $1464 

UV/VIS $300 to $ 504 

AA $228 to $ 360 

797.2800 Early seedling growth toxicity test 

GC/LC $570 to $1220 

UV/VIS $250 to $ 420 

AA $190 to $ 300 

Marine sediment toxicity test - Schwartz 

GC/LC $1368 to $2928 

UV/VIS $ 600 to $1008 

AA $ 456 to $ 720 

10 samples 

24 samples 

12 

J 
P.91 

ATTACHMENT I (CONTINUED) 

HEALTH EFFECTS TESTS 

798.1100 N/A 

798.1150 Acute inhalation toxicity 

4 intervales x 3 concentrations + control -

16 samples 

GC/LC $912 to $1952 

UV/VIS $400 to $ 672 

AA $304 to $ 480 

798.1175 N/A 

798.2250 N/A 

798.2450 Inhalation toxicity 

Initial and weekly monitoring on 3 concentrations and 

contraol at 3 time periods for each interval -

168 samples 

GC/LC $9576 to $20496 

UV/VIS $4200 to $ 7056 

AA $3192 to $ 5040 

798.2650 oral toxicity 

Gavage and drinking water 

Stability assays 24 samples 

Weekly concentration assays 52 samples 

Total 76 samples 

GC/LC $4322 to $9272 

UV/VIS $1900 to $3192 

AA $1444 to $2280 

Dietary 

Same as above + homogeneity of mixing 

24 samples 

Total 100 samples 

GC/LC $5700 to $12200 

UV/VIS $2500 to $ 4200 

AA $1900 to $ 3000 

13 
ATTACHMENT I (CONTINUED) 

HEALTH EFFECTS TESTS 

P.92 

798.3300 

798.4100 

798.4700 

Oncogenicity 

Gavage and drinking water 

Stability assays 

Concentration assasy 

Weekly for 13 weeks followed 

by monthly for 21 months 

Total 

24 samples 

280 

304 

GC/LC 

UV/VIS 

AA 

Dietary 

Same as above + 

$17328 to $37088 

$ 7600 to $12768 

$ 5776 to $ 9120 

homogeneity of mixing 

24 

Total 328 

$18696 to $40016 

$ 8200 to $13776 

$ 6232 to $ 9840 

samples 

samples 

GC/LC 

UV/VIS 

AA 

Inhalation 

Equilibrium assays 

Weekly chamber monitoring 

at 3 intervals per chamber 1248 

Total 1296 

GC/LC $73872 to $258112 

UV/VIS $32400 to $ 54432 

AA $24624 to $ 38880 

samples 

samples 

48 samples 

samples 

samples 

N/A 

Reproduction and fertility effects 

Stability assays 

Concentration assays 

Weekly for 4 weeks followed by 

monthly for 10 months 

Total 

GC/LC 

UV/VIS 

AA 

Dietary 

Same as above 

24 samples 

56 

80 

GC/LC 

UV/VIS 

AA 

$4560 to $9760 

$2000 to $3360 

$1520 to $2400 

homogeneity of mixing 

24 

Total 104 

$5928 to $12688 

$2600 to $ 4368 

$1976 to $ 3120 

samples 

samples 

samples 

samples 

14 
P.93 

ATTACHMENT I (CONTINUED) 

HEALTH EFFECTS TESTS 

798.4700 Reproduction and fertility effects (continued) 

Inhalation 

Equilibration assays 48 samples 

Weekly chamber monitoring 

at 3 intervals per chamber 480 samples 

Total 528 samples 

GC/LC $30096 to $64416 

UV/VIS $13200 to $22176 

AA $10032 to $15840 

798.4900 N/A 

SUBPART F - GENETIC TOXICITY 

Because of the nature of these tests, no analytical 

determination are generally required. Exceptions may include 

those mammalian tests with multiple dose regimes or via the 

inhalation routes. 

SUBPART G - NEUROTOXICITY 

798.6050 Functional observation battery 

798.6200 Motor activity 

798.6400 Neuropathology 

798.6500 Schedule-controlled operant behavior 

Analytical costs are dependent on the exposure 

protocol for the animals on these tests. 

15 
ATTACHMENT II 

DESCRIPTION OF ANALYTICAL CHEMISTRY TECHNIQUES 

UV/VIS: ULTRAVIOLET/VISIBLE ABSORBANCE SPECTROSCOPY 

Instrumental method for measuring the ocncentration of a compound 

in solution based on it's ability to absorb light at a particular 

wavelength. This technique is generally applicable to very 

simple solutions of a few components. It is usually applied to 

the measurement of organic compounds. The test solutions must be 

clear (free of particulate matter), with only the component of 

interest absorbing light at a particular wavelength. Test 

solutions are usually poured into transparent vessels that are 

inserted one at a time into the instrument and the absorbance 

value (amount of light absorbed) is read off a gauge or digital 

display. The major complications in this type of analysis are 

dissolution of the compound in an appropriate solvent and/or 

clarification of the resulting solution. This is one of the 

simpler instrumental methods of analysis. Interferences from 

various components of a complex sample (one with many substances 

other than the compound of interest present) limit usefulness of 

it in many circumstances. Some samples can be analyzed directly 

while others may require some pretreatment. 

LC/GC: LIQUID CHROMATOGRAPHY/GAS CHROMATOGRAPHY 

Instrumental method for measuring the concentration of a compound 

or group of compounds in a solution by separating them along a 

stream flowing liquid or gas which passes continuously through a 

detector. The stream is passed through a column packed with 

material that creates the separation of components before 

reaching the detector. The detector responds to specific 

physical or chemical characteristics of the compounds including 

but by no means limited to light absorption. Separation and 

detection of the compounds usually occurs over a time span from a 

few minutes to several hours depending upon the analysis 

conditions. The choice of such parameters as the type of column, 

detector, liquid or gas moving through the system, temperature 

and many other factors can determine the time for the analysis of 

a solution as well as the amount of sample preparation needed 

before analysis. The benefits of LC/GC analysis over simple 

absorbance spectroscopy are added sensitivity for most compounds, 

the ability to analyze very complex mixtures and more than one 

component in a solution. Solutions must be free of particulates, 

and compounds that are not compatible with the flowing liquid or 

gas. Sample preparation can often involve several steps designed 

to dissolve a compound in a particular solvent, put it into a 

form that gives a response in a detector and/or remove 

interfering substances. The major complications of LC/GC 

16 
P.95 

ATTACHMENT II (CONTINUED) 

DESCRIPTION OF ANALYTICAL CHEMISTRY TECHNIQUES (CONTINUED) 

analysis are instability of instruments, interferences from 

similar compounds and interactions between the flowing streams 

and the columns that produce the separation of components. These 

techniques are usually applied to organic compounds. Most 

samples require some pretreatment. 

AA: ATOMIC ABSORPTION SPECTROPHOTOMETRY 

Instrumental method for measuring the concentration of inorganic 

(elemental) components of a solution based on their ability to 

absorb light at particular wavelengths when heated by a flame or 

electric current. When elements are heated enough to vaporize 

and form atoms, they will absorb light at wavelengths 

characteristic of each element. In this type of analysis of a 

solution containing an element is burned as part of a fule 

mixture or heated by an electrical current. At high enough 

temperatures atoms are formed in the exhaust gases. A beam of 

light from a lamp that emits light at the same wavelengths as 

those absorbed by the element is passed through the vapor. The 

amount of light that is absorbed in the vapor is proportional to 

the concentration of the element in the solution. The element 

being analyzed for determines the type of lamp, wavelength being 

monitored by the instrument, type (temperature) of the flame and 

sensitivity of the assay. There are a number of known 

interferences in AA analysis. Many of them can be eliminated or 

at least minimized by modifications of the technique. The major 

complications of the method are removing organic components from 

a sample without loss of the element of interest, dissolution of 

the element and removal of inter-element interferences. Almost 

all samples analyzed by this method require some form of 

pretreatment raning from simple addition of an acid to extensive 

ashing and dissolution procedures fro very complex samples. 

17 
P.96 

III. RANGE-FINDING TESTS 

A major objective in the toxicological assessment of any 

test chemical is the determination of both clearcut effect and no 

effect levels. These levels are equally important for both 

health effects assessments and environmental impact evaluations. 

In order to select the appropriate dose or concentration ranges 

for the definitive tests, range-finding tests are frequently 

conducted. 

The Toxic Substances Control Act (TSCA) testing guidelines 

for many of the environmental effects tests recognize the 

importance of range-finding evaluations. Although not a 

requirement of any of the test guidelines, statements such as "a 

range-finding test should be conducted" or "it may be necessary 

to perform a range-finding test" are found in all of the aquatic 

test guidelines except for the bioconcentration tests. The 

bioconcentration tests require information from acute toxicity 

tests to establish the appropriate concentration for evaluation. 

In these cases, the acute tests can be considered as rangefinding tests. 

In the examples above, the main purpose of the range-finding 

tests is to establish the test chemical concentrations to be used 

in the definitive tests. In specific cases, such as the Algal 

Acute Toxicity Test (797.1050) and the Lemna Acute Toxicity Test 

(797.1160), the results from the range-finding may determine that 

a definitive test is not necessary. The range-finding test for 

18 
P.97 

the Mysid Shrimp Acute Toxicity Test (797.1930) is also necessary 

to detex-mine which life stage (juvenile or young adult) should be 

utilized in the definitive test. 

The Health Effects Testing Guidelines do not use the terminology "range-finding tests" within the guidelines, however, a 

few studies discuss range-finding tests using other terminology. 

The Inhalation Developmental Toxicity (798.4350), the Preliminary 

Developmental Toxicity Screen (798.4420) and Developmental 

Toxicity (798.4900) guidelines state "a pilot or trial study may 

be advisable" for the selection of the appro-priate dose levels. 

Some of the Genetic Toxicity guidelines for various plate assays 

discuss initially performing the test with a broad range of concentrations. Other tests do not include such statements. 

The majority of the Health Effect tests are specific in the 

number of dose groups that must be used and the criteria for 

acceptability of the study with a clear cut dose response 

relationship and a no-observed-toxic-effect level. The highest 

dose level in rodents should result in toxic effects, but not 

produce an incidence of fatalities which would prevent a 

meaningful evaluation. The lowest dose level should not produce 

any evidence of toxicity. In order to meet these criteria, it is 

important to consider well designed, test chemical specific 

range-finding tests. 

19 
P.98 

The progression in the development of toxicology programs 

includes the completion of the acute tests and repeated dose 

range-finding tests to select doses for the subchronic (SC), 

developmental (DEV) and reproductive (REP) tests, the conduct of 

the SC, DEV and REP, as required, and, if necessary, the conduct 

of the chronic and oncogenicity tests. The results of all previous tests are used in the selection of the dose to be used in 

the chronic tests. In cases where no previous data exist in a 

particular animal species by a specific dosing regimen, further 

range-finding tests may be conducted. As an example, the majority of the acute, subchronic and reproductive tests are conducted 

in the rat. This data would not be meaningful in the selection 

of doses to be used in a mouse chronic or oncogenicity test. 

Therefore, additional "range-finding" studies appropriate to that 

species may be necessary. 

The study design for range-finding should follow the same 

principles and practices of the definitive study. In the rangefinding test a limited number of the species under test are 

exposed to a wide range test chemical concentrations or doses for 

a duration considered to be acceptable to ascertain results that 

are meaningful to the definitive study. For example, a 4-6 week 

range-finding study with five mice per sex per group dosed via 

the diet at 5 or 6 broadly spaced concentrations may be 

sufficient to select the levels to be tested in a 2-year mouse 

oncogenicity by the dietary route. 

20 
P.99 

The range-finding tests recommended for the Environmental 

Effects guidelines are outlined in sufficient detail so that one 

can effectively plan the test. The Health Effects tests, 

however, require the skills of the toxicologist in reviewing the 

purpose of the definitive test and ascertaining the potential 

toxicities of the test chemical so that the range-finding can be 

properly designed. As an example, the acute oral LD50 of the 

chemical has been determined and it is known that single oral 

doses cause moderate to severe irritation of the stomach. The 

range finding test for a chronic oral gavage study would be 

designed to find dose ranges that would neither be toxic nor 

produce stomach irritation upon repeated administration. As a 

general rule, range-finding data should be generated prior to the 

initiation of any definitive test in which the endpoints are the 

determination of clearcut effect and no effect levels or where 

additional information is necessary before the definitive test 

can initiate, e.g., a non-cytotoxic level for a mammalian cell 

culture test. 

Because of the lack of definition for range-finding tests in 

Health Effects and the uncertainty of conducting range-finding 

tests in the Environmental Effects guidelines, the costs 

associated with these tests are not included in the test cost 

evaluations. Some consideration should be given to including 

these costs in the future. 

21 
P.1 

VINCENT J. PICCIRILLO 

TECHNICAL QUALIFICATIONS 

o 

o 

Board-certified toxicologist with ten years experience in 

chemical risk assesssment. 

Leading expert in TSCA testing requirements. 

Seven years experience with OTS regulatory analysis. 

EDUCATION; 

1976 

1974 

1970 

Diplomate, American Board of Toxicology 

Ph.D., Medical College of Virginia, Pharmacology 

B.A., Pennsylvania State University, Biological 

Science 

1967-78 

1974-76 

Consultant, Mathtech, Inc. 

Director of Toxicology, Tegeris Laboratories, Inc. 

(formerly Borriston Laboratories, Inc.) 

Senior Staff Scientist, Hazelton Laboratories 

Senior Toxicologist, Rohm and Haas 

EXPERIENCE: 

As an independent consultant, Dr. Piccirillo assists Mathtech, 

Inc. in developing protocols for toxicological and environmental 

tests required by U.S.EPA under Sections 4 and 5 of the Toxic 

Substances Control Act. He has also assisted in the development 

of an automated model to determine the costs of such tests and in 

the preparation of the TSCA Test Cost Manual. 

As toxicology division director for Tegeris Laboratories, Dr. 

Piccirillo was responsible for financial, technical and 

managerial aspects of all studies conducted by the division. The 

division was comprised of approximately 20 senior and junior 

scientists, technicians, report writers/data coordinators and 

animal caretakers with an annual budget in excess of $2 million. 

He worked closely with industrial sponsors in assuring that study 

protocols complied with intended registration guidelines or other 

test standards (FIFRA, TSCA, OECD, FDA, OSHA). Technical 

Page 1 of 4 
P.2 

expertise covered the design, implementation, evaluation and 

report preparation of acute, subchronic and chronic/oncogenic 

studies in rodents, rabbits, guinea pigs and canines by all 

classical routes of administration (intubation, feeding, dermal, 

ocular, intravenous, water dosing, etc.). He served as principal 

investigator for two major NIOSH contracts to evaluate the 

Chernoff techniques as an effective screen for reproductive 

hazard potential of priority chemicals. 

Also with Tegeris, Dr. Piccarillo served as the program manager 

for a subcontract to perform cost and economic analysis of TSCA, 

FIFRA, and OECD test standards and test rules for EPA. This 

program vas expanded to include evaluation of test standards 

under the Premanufacture Notification Program (PMN) of TSCA. He 

was laboratory representative to the Regulatory Affairs Committee 

of the National Association for Life Science Industries (NALSI), 

where he was involved in review, comment and dissemination of 

information regarding regulatory matters affecting member 

companies. 

With Hazelton Laboratories, Dr. Piccarillo conducted research 

studies to evaluate the safety of various test materials 

including drugs, cosmetics, food and color additives, and 

industrial, agricultural and household chemicals in experimental 

animals. He was responsible for the development of experimental 

study designs, coordination of the laboratory work, and 

interpretation of the data resulting from acute, subacute and 

chronic toxicity studies as well as teratology, carcinogencity 

and reproduction studies. Other activities included supervision 

of the canine toxicology laboratory. 

While with Rohm and Haas, Dr. Piccarillo was responsible for 

establishing in-house subchronic and specialized toxicity testing 

capabilities. He was also responsible for designing protocols, 

monitoring projects, supervising personnel, and writing reports. 

In addition, he monitored toxicity studies awarded to extramural 

research testing laboratories and prepared Section C submissions 

to the Environmental Protection Agency covering herbicide and 

plant fungicide areas. 

AFFILIATIONS; 

Society of Toxicology 

American College of Toxicology 

American Association for Laboratory Animal Science (AALAS) 

National Association of Life Science Industries (NALSI), 

Regulatory Affairs Committee 

Page 2 of 4 
P.3 

PUBLICATIONS AND PRESENTATIONS: 

Chambers, J.W. and Piccirillo, V.J., The effects of ethanol on 

amino acid uptake and utilization by the liver and other 

organs, Quart. J. stud. Ale. 14: (3), 707-719. (1973) 

Piccirillo, V.J. (Doctoral Thesis). The effects of ethanol 

on amino acid metabolism in the liver and other organs in 

the rat. Medical College of Virginia. (1974) 

Piccirillo, V.J. and Chambers, J.W., The effects of ethanol 

metabolites on urea production by the isolated perfused rat 

liver. IRCS Med. Sci. 3: 186. (1975) 

Piccirillo, V.J. and Chambers, J.W., Inhibition of hepatic 

uptake of alpha aminoisobutryic acid by ethanol: Effect of 

of pyrazole and metabolites of ethanol. Res. Commun. Chem. 

Path. Pharmcaol. 12: (2) 297. (1976). 

Piccirillo, V. J. and Quesenberry, R.P., Reproductive capacities 

of control mallard ducks (Anas Platyrhynchos) during a one 

generation reproduction study, J. Env. Path. Toxicol. 4, 

Parts 5. 6: 133-140. (1980). 

Piccirillo, V.J. and Owen, T.B., Inhalation Bioassay of Cigarette 

Smoke in Dogs. Directory of on-going research in Smoking 

and Health. (1980) 

Piccirillo, V.J., Orlando, D.A., and Quesenberry, R.P., 

Reproductive capacities of control bobwhite quail (Colinus 

virginianus) during a one generation reproduction study. J 

Env. Path. Toxicol. (Accepted January, 1981) 

Parker, CM., Piccirillo, V.J., Kurtz, S.L., Garner, F.M., 

Gardiner, T.H. and Van Gelder, G.A. Six month feeding study 

of Fenvalerate in Dogs. Fundam. Appl. Toxicol. 4(4): 

577-586 (1984). 

Schuler, R.L., Hardin, B.D., Neimeier, R.W., Booth G., Hazleden, 

K., Piccirillo, V.J. and Smith, K. Results of testing 

fifteen glycol ethers in a shortterm in vivo reproductive 

toxicity assay. Environ. Health Perspect. 57:141-146 

(1984). 

Brown, J.H., Piccirillo, V.J. and Hartman, W.C, Jojobutter-51, A 

primary dermal irritation study in rabbits. Cosmetics and 

toiletries, 99(51:57 (1984). 

Haider, C.A., Holdsworth, C.E., Cockrell, B.Y., and Piccirillo, 

V.J., Hydrocarbon nephropathy in male rats: Identification 

of the nphrotoxic components of unleaded gasoline. Tox. and 

Industr. Health. (Accepted September, 1985). 

Page 3 of 4 
P.4 

Piccirillo, V.J., and Chambers, J.W., The effect of pyrazole on 

the inhibition of AIB uptake by ethanol in the isolated 

perfused rat liver. Fed Proc. 32: 698. (1973). Presented 

at the FASEB meeting, Atlantic City, New Jersey, 1973. 

Chambers, J. W. and Piccirillo, V.J., Increase in amino acid 

incorporation into protein and decrease in' hepatic urea 

synthesis produced by ethanol. Fed. Proc. 3_3_: 554, 

(1974). Presented at the FASEB meeting, Atlantic City, New 

Jersey, 1974. 

Piccirillo, V.J. and Chambers, J.W. Ethanol effect on 

incorporation of alanine into protein in various organs of 

the rat. The Pharmacologis 16.: (2). 303, (1974). 

Presented at the ASPET meeting Montreal, Canada, 1974. 

Piccirillo, J.V. and Quesenberry, R.P., Biological variation in 

the reproductive capacities of control mallard ducks (Anas 

platyrhynchos) during one generation reproduction studies. 

Toxicol. Appl. Pharmacol. 4J3.: Part 2: 1979. Presented at 

the SOT meeting, New Orleans, Louisiana, 1979. 

Parker, CM., Piccirillo, V.J., Kurtz, S.L., and Garner, F.M., 

Six Month Feeding Study of Fenvalerate in Dogs. The 

Toxicologist, Vo., No. 1, March, 1983. Presented at the SOT 

meeting, Las Vegas, Nevada, 1983. 

Neal, B.H., Piccirillo, V.J., Rowland, J.C and Kastello, M.D., 

Preclinical Toxicity of Vehicle 12 in Beagle Dogs. The 

Toxicologist, Vo. 3, No. 1, March, 1983. Presented at the 

SOT meeting, Las Veags, Nevada, 1983. 

Lunchick, C , Hartman, W.C. and Piccirillo, V.J. Variability of 

the dominant lethal response in the rat to orally 

administered triethylenemelamine. Presented at the NCAB 

chapter of SOT meeting. Spring, 1983. 

Piccirillo, V.J., Health Effects and Environment Effects 

Guidelines: Evaluation and Cost Estimation. Presented to 

the Regulatory Impacts Branch, U.S. Environmental Protection 

Agency, Washington, D.C. on August 6 and 13, 1985. 

Piccirillo, V.J., J.K. Orrell, and P.M. Godfrey, TSCA Test 

Guideline Cost Estimates prepared for the Economics and 

Technology Division, Office of Toxic Substances, U.S. 

Environmental Protection Agency, Washington, D.C. 

Page 4 of 4